def makePreMix(preMix, NumSamples=None):
NumSamples = NumSamples or Rtv.NumOfSamples
l = preMix.labware
msg = "preMix: {:.1f} µL of {:s}".format(preMix.minVol(NumSamples),
preMix.name)
with group(msg):
msg += " into {:s}[grid:{:d} site:{:d} well:{:d}] from {:d} components:".format(
l.label, l.location.grid, l.location.site + 1, preMix.pos + 1,
len(preMix.components))
Itr.comment(msg).exec()
nc = len(preMix.components)
nr = len(preMix.Replicas)
nt = Rbt.Robot.current.curArm().nTips
assert nc <= nt, "Temporally the mix can not contain more than {:d} components.".format(
nt)
dt = nt - nc
samples_per_replicas = [(NumSamples + nr - (i + 1)) // nr
for i in range(nr)]
with tips(Rbt.tipsMask[nc]): # want to use preserved ?? selected=??
for i, react in enumerate(preMix.components):
l = react.labware
r = react.volpersample * NumSamples * preMix.excess
msg = " {:d}- {:.1f} µL of {:s} from {:s}[grid:{:d} site:{:d} well:{:d}]".format(
i + 1, r, react.name, l.label, l.location.grid,
l.location.site + 1, react.pos + 1)
Itr.comment(msg).exec()
mV = Rbt.Robot.current.curArm(
).Tips[i].type.maxVol # todo what if the tip are different?
while r > 0:
dV = r if r < mV else mV
aspire(i, react, dV)
multidispense_in_replicas(
i, preMix,
[sp / NumSamples * dV for sp in samples_per_replicas])
r -= dV
def Run(self):
self.set_EvoMode() # this add: self.iRobot = EvoMode.iRobot(Itr.Pipette.LiHa1, nTips=self.nTips)
# in which: self.robot = Rbt.Robot(index=index, arms=arms, nTips=nTips)
self.initialize() # if needed calls Executable.initialize() and set_EvoMode
# which calls GUI.update_parameters() and set_defaults() from Evo200
Rtv.NumOfSamples = self.NumOfSamples
NumOfSamples = self.NumOfSamples
wt = self.worktable
Itr.comment('Prefill {:d} plates with LysisBufferReact for {:d} samples.'\
.format(self.num_plates,
NumOfSamples )).exec()
# Get Labwares (Cuvette, eppys, etc.) from the work table
BufCuvette = wt.getLabware(Lab.Trough_100ml, "2-Vl Lysis Buffer")
self.go_first_pos() # Set the initial position of the tips
# Set volumen / sample
BufferVolume = 100.0 # VL1 or VL
all_samples = range(NumOfSamples)
maxTips = min (self.nTips, NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
# Define the reactives in each labware (Cuvette, eppys, etc.)
BufferReact = Rtv.Reagent("Buffer ",
BufCuvette,
volpersample = BufferVolume,
defLiqClass = 'MN VL',
num_of_samples= self.num_plates * NumOfSamples)
# Show the CheckList GUI to the user for possible small changes
self.CheckList()
self.set_EvoMode()
Itr.wash_tips(wasteVol=5, FastWash=True).exec()
LysPlat = [wt.getLabware(Lab.MP96deepwell, "Plate lysis-"+str(i+1)) for i in range(self.num_plates)]
par = LysPlat[0].parallelOrder(self.nTips, all_samples)
# Define place for temporal reactions
for i, LP in enumerate(LysPlat):
for s in all_samples:
Rtv.Reagent("lysis_{:d}-{:02d}".format(i + 1, s + 1),
LP,
initial_vol =0.0,
pos =s + 1,
excess =0)
with group("Prefill plates with BufferReact"):
Itr.userPrompt("Put the plates for BufferReact").exec()
for LP in LysPlat:
with self.tips(reuse=True, drop=False):
self.spread(reactive=BufferReact, to_labware_region=LP.selectOnly(all_samples))
self.dropTips()
self.done()
def Run(self):
self.set_EvoMode()
self.initialize() # set_defaults ??
Rtv.NumOfSamples = self.NumOfSamples
NumOfSamples = self.NumOfSamples
wt = self.worktable
Itr.comment('Prefill plates with VEW1, Elution buffer and VEW2 for {:s} samples.'.format(str(NumOfSamples))).exec()
# Get Labwares (Cuvette, eppys, etc.) from the work table
ElutBuf = wt.getLabware(Lab.Trough_100ml, "1-VEL-ElutionBuffer" )
DiTi1000_1 = wt.getLabware(Lab.DiTi_1000ul, "1000-1")
DiTi1000_2 = wt.getLabware(Lab.DiTi_1000ul, "1000-2")
DiTi1000_3 = wt.getLabware(Lab.DiTi_1000ul, "1000-3")
Plate_VEW1 = wt.getLabware(Lab.MP96deepwell, "Plate VEW1" ) # Plate 12 x 8 ?
Plate_VEW2 = wt.getLabware(Lab.MP96deepwell, "Plate VEW2" ) # Plate 12 x 8 ?
Plate_Eluat = wt.getLabware(Lab.MP96well, "Plate ElutB" ) # Plate 12 x 8 ? MP96well !!
# Set the initial position of the tips
self.go_first_pos()
# Set volumen / sample
VEW1Volume = 600.0
VEW2Volume = 600.0
ElutionBufferVolume = 100.0
# Liquid classes used for pippetting. Others liquidClass names are defined in "protocol_steps.py"
# SampleLiqClass = "Serum Asp" # = TissueHomLiqClass # SerumLiqClass="Serum Asp preMix3"
all_samples = range(NumOfSamples)
maxTips = min (self.nTips, NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
# Define the reactives in each labware (Cuvette, eppys, etc.)
VEW1 = Rtv.Reagent("VEW1 - Wash Buffer ",
wt.getLabware(Lab.Trough_100ml, "4-VEW1 Wash Buffe"),
volpersample = VEW1Volume,
defLiqClass = B_liquidClass)
VEW2 = Rtv.Reagent("VEW2 - WashBuffer ",
wt.getLabware(Lab.Trough_100ml, "5-VEW2-WashBuffer" ),
volpersample =VEW2Volume,
defLiqClass =B_liquidClass)
ElutionBuffer = Rtv.Reagent("Elution Buffer ",
ElutBuf,
volpersample =ElutionBufferVolume,
defLiqClass =B_liquidClass)
# Show the CheckList GUI to the user for posible small changes
self.CheckList()
self.set_EvoMode()
Itr.wash_tips(wasteVol=30, FastWash=True).exec()
par = Plate_VEW1.parallelOrder(self.nTips, all_samples)
# Define samples and the place for temporal reactions
for s in all_samples:
Rtv.Reagent("VEW1_{:02d}".format(s + 1),
Plate_VEW1,
initial_vol = 0.0,
pos = par[s]+1,
excess = 0) # todo revise order !!!
Rtv.Reagent("VEW2_{:02d}".format(s + 1),
Plate_VEW2,
initial_vol = 0.0,
pos = par[s] + 1,
excess = 0)
Rtv.Reagent("Eluat_{:02d}".format(s + 1),
Plate_Eluat,
initial_vol = 0.0,
pos = par[s] + 1,
excess = 0)
with group("Prefill plates with VEW1, Elution buffer and VEW2"):
Itr.userPrompt("Put the plates for VEW1, Elution buffer and VEW2 in that order").exec()
with self.tips(reuse=True, drop=False):
self.spread(reactive=ElutionBuffer, to_labware_region=Plate_Eluat.selectOnly(all_samples) ) # ,optimize=False
with self.tips(reuse=True, drop=False):
self.spread(reactive=VEW2, to_labware_region=Plate_VEW2.selectOnly(all_samples) ) # , optimize=False
with self.tips(reuse=True, drop=False):
self.spread(reactive=VEW1, to_labware_region=Plate_VEW1.selectOnly(all_samples)) # , optimize=False
self.dropTips()
self.done()
def Run(self):
self.set_EvoMode()
self.initialize() # set_defaults ??
Rtv.NumOfSamples = self.NumOfSamples
NumOfSamples = self.NumOfSamples
wt = self.worktable
Itr.comment('Extracting RNA from {:s} samples with the MN-Vet kit'.format(str(NumOfSamples))).exec()
# Get Labwares (Cuvette, eppys, etc.) from the work table
LysBuf = wt.getLabware(Lab.Trough_100ml, "2-Vl Lysis Buffer" )
BindBuf = wt.getLabware(Lab.Trough_100ml, "3-VEB Binding Buffer" )
DiTi1000_1 = wt.getLabware(Lab.DiTi_1000ul, "1000-1")
DiTi1000_2 = wt.getLabware(Lab.DiTi_1000ul, "1000-2")
DiTi1000_3 = wt.getLabware(Lab.DiTi_1000ul, "1000-3")
Reactives = wt.getLabware(Lab.GreinRack16_2mL, "Reactives" )
# Set the initial position of the tips
self.go_first_pos()
# Set volumen / sample
ProtKVolume = 20.0
cRNAVolume = 4.0
LysisBufferVolume = 100.0 # VL1 or VL
IC_MS2Volume = 10.0 # IC2
IC2Volume = 5.0 # ? 4
BindingBufferVolume = 350.0 # VEB
B_BeadsVolume = 20.0 # B-Beads
EtOH80pVolume = 600.0
SampleVolume = 100.0
InitLysisVol = 0.0
if self.version == 'in-VL inactivated':
InitLysisVol = SampleVolume + LysisBufferVolume
elif self.version == 'pre Inactivated':
InitLysisVol = SampleVolume + ProtKVolume + cRNAVolume + IC_MS2Volume + LysisBufferVolume
# Liquid classes used for pippetting.
# Others liquidClass names are defined in "protocol_steps.py"
SampleLiqClass = "Serum Asp" # = TissueHomLiqClass # SerumLiqClass="Serum Asp preMix3"
all_samples = range(NumOfSamples)
maxTips = min (self.nTips, NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
# Define the reactives in each labware (Cuvette, eppys, etc.)
if self.version != 'pre Inactivated': # we need to add ProtK+cRNA+MS2 mix
ProtK = Rtv.Reagent("Proteinase K ",
Reactives,
replicas = 2,
pos = [15, 16],
volpersample = ProtKVolume,
defLiqClass = Small_vol_disp)
cRNA = Rtv.Reagent("Carrier RNA ", Reactives, pos=14, volpersample= cRNAVolume, defLiqClass=Small_vol_disp)
IC_MS2 = Rtv.Reagent("IC MS2 phage culture ", Reactives, pos=13, volpersample= IC_MS2Volume, defLiqClass=Small_vol_disp)
# IC2 = Rtv.Reagent("IC2 - synthetic RNA " , Reactives, pos=13, volpersample= IC2Volume ,defLiqClass=W_liquidClass)
pK_cRNA_MS2 = Rtv.preMix ("ProtK+cRNA+IC-MS2 mix " ,
Reactives,
pos=8,
components=[ cRNA, ProtK, IC_MS2] ,
defLiqClass=W_liquidClass,
excess=20)
if self.version != 'in-VL inactivated':
LysisBuffer = Rtv.Reagent("VL - Lysis Buffer ", LysBuf, volpersample=LysisBufferVolume, defLiqClass='MN VL')
B_Beads = Rtv.Reagent("B - Beads ",
Reactives,
pos = [1,2],
initial_vol = 1200,
volpersample = B_BeadsVolume,
defLiqClass = Beads_LC_2,
maxFull = 70)
VEB = Rtv.Reagent("VEB - Binding Buffer ", BindBuf, volpersample=BindingBufferVolume, defLiqClass=B_liquidClass)
EtOH80p = Rtv.Reagent("Ethanol 80% ", wt.getLabware(Lab.Trough_100ml, "7-EtOH80p"), volpersample=EtOH80pVolume, defLiqClass=B_liquidClass)
# Show the CheckList GUI to the user for possible small changes
self.CheckList()
self.set_EvoMode()
# Define the reactives not shown in the CheckList GUI
# Define samples and the place for temporal reactions
Plate_lysis = wt.getLabware(Lab.MP96deepwell, "Plate lysis" ) # Plate 12 x 8 ?
Plate_EtOH = wt.getLabware(Lab.MP96deepwell, "Plate EtOH" ) # Plate 12 x 8 ? MP96well !!
if self.version != 'original samples':
Samples = wt.getLabware(Lab.EppRack6x16, "Proben" ) # 6x16 = 12 x 8 ?
par = Plate_lysis.parallelOrder(self.nTips, all_samples)
for s in all_samples:
if self.version == 'original samples':
Rtv.Reagent("probe_{:02d}".format(s + 1), Samples, single_use=SampleVolume,
pos=s + 1, defLiqClass=SampleLiqClass, excess=0)
Rtv.Reagent("lysis_{:02d}".format(s + 1), Plate_lysis, initial_vol=InitLysisVol, pos=s + 1,
excess=0)
Rtv.Reagent("EtOH80p_{:02d}".format(s + 1), Plate_EtOH, initial_vol=0.0, pos=par[s] + 1,
excess=0) # todo revise order !!!
Itr.wash_tips(wasteVol=30, FastWash=True).exec()
with group("Prefill plate with EtOH80p"):
with self.tips(reuse=True, drop=False, drop_last=True):
self.spread(reactive=EtOH80p, to_labware_region=Plate_EtOH.selectOnly(all_samples))
with group("Sample Lysis"):
if self.version != 'pre Inactivated': # add ProtK+cRNA+MS2 mix
with self.tips(tipsMask=maxMask, reuse=True, drop=False, drop_last=True):
self.makePreMix(pK_cRNA_MS2)
self.spread ( reactive=pK_cRNA_MS2, to_labware_region= Plate_lysis.selectOnly(all_samples))
if self.version != 'in-VL inactivated': # add LysisBuffer
with self.tips(tipsMask=maxMask, reuse=True, drop=False, drop_last=True):
self.spread ( reactive=LysisBuffer, to_labware_region= Plate_lysis.selectOnly(all_samples))
if self.version == 'original samples': # add samples
Itr.userPrompt("Please make sure the samples are in place").exec()
with self.tips(reuse=False, drop=True):
self.transfer( from_labware_region= Samples,
to_labware_region= Plate_lysis,
volume= SampleVolume,
using_liquid_class= (SampleLiqClass, "Serum Disp postMix3"),
optimizeFrom = False,
optimizeTo = False, # todo Really ??
NumSamples = NumOfSamples)
Itr.wash_tips(wasteVol=4, FastWash=True).exec()
if self.version != 'pre Inactivated':
Itr.userPrompt("Please Schutteln the plates for lysis in pos 1").exec()
with incubation(minutes=5): pass
if self.version != 'prefill inactivation':
Itr.userPrompt("Please make sure the samples are back in place").exec()
with group("Beads binding"):
with self.tips(tipsMask=maxMask, reuse=True, drop=False):
for p in [40, 50, 60, 65]:
self.mix_reactive(B_Beads, LiqClass=Beads_LC_1, cycles=1, maxTips=maxTips, v_perc=p)
with self.tips(reuse=True, drop=False):
self.spread( reactive=B_Beads, to_labware_region=Plate_lysis.selectOnly(all_samples))
self.dropTips()
with self.tips(reuse=True, drop=False):
self.spread( reactive=VEB, to_labware_region=Plate_lysis.selectOnly(all_samples))
Itr.userPrompt("Please Schutteln the plates for lysis in pos 1").exec()
self.dropTips()
self.done()
def Run(self):
self.set_EvoMode()
self.initialize()
Rtv.NumOfSamples = self.NumOfSamples
NumOfSamples = self.NumOfSamples
wt = self.worktable
Itr.comment('Prefill {:d} plates with LysisBufferReact for {:d} samples.'\
.format(self.num_plates,
NumOfSamples )).exec()
# Get Labwares (Cuvette, eppys, etc.) from the work table
LysBufCuvette = wt.getLabware(Lab.Trough_100ml, "2-Vl Lysis Buffer")
DiTi1000_1 = wt.getLabware(Lab.DiTi_1000ul, "1000-1")
DiTi1000_2 = wt.getLabware(Lab.DiTi_1000ul, "1000-2")
DiTi1000_3 = wt.getLabware(Lab.DiTi_1000ul, "1000-3")
self.go_first_pos() # Set the initial position of the tips
# Set volumen / sample
LysisBufferVolume = 100.0 # VL1 or VL
all_samples = range(NumOfSamples)
maxTips = min (self.nTips, NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
# Define the reactives in each labware (Cuvette, eppys, etc.)
LysisBufferReact = Rtv.Reagent("VL - Lysis Buffer ",
LysBufCuvette,
volpersample = LysisBufferVolume,
defLiqClass = 'MN VL',
num_of_samples= self.num_plates * NumOfSamples)
# Show the CheckList GUI to the user for possible small changes
self.CheckList()
self.set_EvoMode()
Itr.wash_tips(wasteVol=5, FastWash=True).exec()
LysPlat = [wt.getLabware(Lab.MP96deepwell, "Plate lysis-"+str(i+1)) for i in range(self.num_plates)]
par = LysPlat[0].parallelOrder(self.nTips, all_samples)
# Define place for temporal reactions
for i, LP in enumerate(LysPlat):
for s in all_samples:
Rtv.Reagent("lysis_{:d}-{:02d}".format(i + 1, s + 1),
LP,
initial_vol =0.0,
pos =s + 1,
excess =0)
with group("Prefill plates with LysisBufferReact"):
Itr.userPrompt("Put the plates for LysisBufferReact").exec()
for LP in LysPlat:
with self.tips(reuse=True, drop=False):
self.spread(reactive=LysisBufferReact, to_labware_region=LP.selectOnly(all_samples))
self.dropTips()
self.done()
def mix(self, in_labware_region, using_liquid_class=None, volume=None, optimize=True):
"""
Mix each of the reactive in the selected region of the labware
:param in_labware_region:
:param using_liquid_class:
:param volume:
:param optimize:
:return:
"""
mix_p = 0.9
in_labware_region = in_labware_region or self.worktable.def_WashWaste # todo ???????????
assert isinstance(in_labware_region, Lab.Labware), 'A Labware expected in in_labware_region to be mixed'
if not volume or volume< 0.0 : volume = 0.0
assert isinstance(volume, (int, float))
oriSel = in_labware_region.selected()
nt = self.robot.curArm().nTips # the number of tips to be used in each cycle of pippeting
if not oriSel:
oriSel = range(Rtv.NumOfSamples)
if optimize:
oriSel = in_labware_region.parallelOrder( nt, oriSel)
NumSamples = len(oriSel)
SampleCnt = NumSamples
if nt > SampleCnt:
nt = SampleCnt
# mV = Rbt.Robot.current.curArm().Tips[0].type.maxVol * 0.8
mV = self.worktable.def_DiTi.maxVol * mix_p # What tip tp use !
if volume:
v = volume
else:
v = in_labware_region.Wells[oriSel[0]].vol
v = v * mix_p
v = v if v < mV else mV
lf = in_labware_region
mx = Itr.mix(Rbt.tipsMask[nt], using_liquid_class, volume, in_labware_region)
msg = "Mix: {v:.1f} µL of {n:s}".format(v=v, n=lf.label)
with group(msg):
msg += " [grid:{fg:d} site:{fs:d}] in order:".format(fg=lf.location.grid, fs=lf.location.site+1) \
+ str([i+1 for i in oriSel])
Itr.comment(msg).exec()
while SampleCnt:
curSample = NumSamples - SampleCnt
if nt > SampleCnt:
nt = SampleCnt
mx.tipMask = Rbt.tipsMask[nt]
sel = oriSel[curSample:curSample + nt]
spl = range(curSample, curSample + nt)
with self.tips(Rbt.tipsMask[nt], selected_samples=spl):
mV = self.robot.curArm().Tips[0].type.maxVol * mix_p
mx.labware.selectOnly(sel)
if not using_liquid_class:
if sel:
mx.liquidClass = mx.labware.selected_wells()[0].reactive.defLiqClass
if volume:
r = volume # r: Waste_available yet; volume: to be Waste
else:
vols = [w.vol for w in mx.labware.selected_wells()]
r_min, r_max = min(vols), max(vols)
assert r_min == r_max
r = r_max
r = r * mix_p
r = r if r < mV else mV
mx.volume = r
mx.exec()
SampleCnt -= nt
mx.labware.selectOnly(oriSel)
return oriSel
def waste(self, from_labware_region=None, using_liquid_class=None, volume=None, to_waste_labware=None, optimize=True):
"""
:param from_labware_region:
:param using_liquid_class:
:param volume:
:param to_waste_labware:
:param optimize:
:return:
"""
to_waste_labware = to_waste_labware or self.worktable.def_WashWaste
assert isinstance(from_labware_region, Lab.Labware), 'A Labware expected in from_labware_region to transfer'
if not volume or volume< 0.0 : volume = 0.0
assert isinstance(volume, (int, float))
oriSel = from_labware_region.selected()
nt = self.robot.curArm().nTips # the number of tips to be used in each cycle of pippeting
if not oriSel:
oriSel = range(Rtv.NumOfSamples)
if optimize:
oriSel = from_labware_region.parallelOrder(nt, oriSel)
NumSamples = len(oriSel)
SampleCnt = NumSamples
if nt > SampleCnt:
nt = SampleCnt
tm = Rbt.tipsMask[nt]
nt = to_waste_labware.autoselect(maxTips=nt)
mV = self.worktable.def_DiTi.maxVol # todo revise !! What tip tp use !
Rest = 50 # the volume we cannot more aspire with liquid detection, to small, collisions
RestPlus = 50
CtrVol = 0.5
if volume:
v = volume
else:
v = from_labware_region.Wells[oriSel[0]].vol
Asp = Itr.aspirate(tm, Te_Mag_LC, volume, from_labware_region)
# Asp = Itr.aspirate(tm, using_liquid_class[0], volume, from_labware_region)
Dst = Itr.dispense(tm, using_liquid_class, volume, to_waste_labware)
# Ctr = Itr.moveLiha(Itr.moveLiha.y_move, Itr.moveLiha.z_start, 3.0, 2.0, tm, from_labware_region)
lf = from_labware_region
msg = "Waste: {v:.1f} µL of {n:s}".format(v=v, n=lf.label)
with group(msg):
msg += " [grid:{fg:d} site:{fs:d}] in order:".format(fg=lf.location.grid, fs=lf.location.site+1) \
+ str([i+1 for i in oriSel])
Itr.comment(msg).exec()
while SampleCnt:
curSample = NumSamples - SampleCnt
if nt > SampleCnt:
nt = SampleCnt
tm = Rbt.tipsMask[nt]
Asp.tipMask = tm
Dst.tipMask = tm
sel = oriSel[curSample:curSample + nt]
spl = range(curSample, curSample + nt)
Asp.labware.selectOnly(sel)
if volume:
r = volume # r: Waste_available yet; volume: to be Waste
else:
vols = [w.vol for w in Asp.labware.selected_wells()]
r_min, r_max = min(vols), max(vols)
assert r_min == r_max
r = r_max
if not using_liquid_class:
if sel:
Dst.liquidClass = Asp.labware.selected_wells()[0].reactive.defLiqClass
with self.tips(tm, drop=True, preserve=False, selected_samples=spl):
while r > Rest: # dont aspire Rest with these Liq Class (Liq Detect)
dV = r if r < mV else mV
if dV < Rest: break # ??
dV -= Rest # the last Rest uL have to be aspired with the other Liq Class
Asp.volume = dV # with Liq Class with Detect: ">> AVR-Serum 1000 << 365"
Dst.volume = dV
Asp.liquidClass = Te_Mag_LC # ">> AVR-Serum 1000 << 365" # "No Liq Detect"
Asp.exec()
Asp.volume = CtrVol
Asp.liquidClass = Te_Mag_Centre
with self.tips(allow_air=CtrVol):
Asp.exec()
if dV + Rest + RestPlus + 2*CtrVol > mV:
Dst.exec()
r -= dV
Dst.volume = 0
else:
break
Asp.volume = Rest
Asp.liquidClass = Te_Mag_Rest # ">> AVR-Serum 1000 << 367" # "No Liq Detect"
with self.tips(allow_air=Rest):
Asp.exec()
Asp.volume = CtrVol
Asp.liquidClass = Te_Mag_Force_Centre
with self.tips(allow_air=CtrVol):
Asp.exec()
Asp.volume = RestPlus
Asp.liquidClass = Te_Mag_RestPlus # ">> AVR-Serum 1000 << 369" # "No Liq Detect"
with self.tips(allow_air=RestPlus):
Asp.exec()
#Ctr.exec()
Asp.volume = CtrVol
Asp.liquidClass = Te_Mag_Force_Centre
Dst.volume += Rest + RestPlus
with self.tips(allow_air=CtrVol):
Asp.exec()
with self.tips(allow_air=Rest + RestPlus):
Dst.exec()
SampleCnt -= nt
Asp.labware.selectOnly(oriSel)
Itr.wash_tips(wasteVol=4).exec()
return oriSel
def Run(self):
self.set_EvoMode()
self.initialize()
Rtv.NumOfSamples = self.NumOfSamples
NumOfSamples = self.NumOfSamples
wt = self.worktable
Itr.comment((self.version + 'for extracting RNA from {:s} samples with the MN-Vet kit').format(str(NumOfSamples))).exec()
# Get Labwares (Cuvette, eppys, etc.) from the work table
if self.add_VL:
LysBuf = wt.getLabware(Lab.Trough_100ml, "2-Vl Lysis Buffer" )
if self.do_extraction:
BindBuf = wt.getLabware(Lab.Trough_100ml, "3-VEB Binding Buffer" )
ElutBuf = wt.getLabware(Lab.Trough_100ml, "1-VEL-ElutionBuffer" )
Eluat = wt.getLabware(Lab.EppRack3x16R, "Eluat")
Samples = wt.getLabware(Lab.EppRack3x16, "Proben")
Lysis = Samples
DiTi1000_1 = wt.getLabware(Lab.DiTi_1000ul, "1000-1")
DiTi1000_2 = wt.getLabware(Lab.DiTi_1000ul, "1000-2")
DiTi1000_3 = wt.getLabware(Lab.DiTi_1000ul, "1000-3")
Reactives = wt.getLabware(Lab.GreinRack16_2mL, "Reactives" )
if self.do_extraction:
self.TeMg_Heat = wt.getLabware(Lab.TeMag48, "48 Pos Heat")
self.TeMag = wt.getLabware(Lab.TeMag48, "48PosMagnet")
TeMag = self.TeMag
Lysis = TeMag
# Set the initial position of the tips
self.go_first_pos()
# Set volumen / sample
SampleVolume = 200.0
LysisBufferVolume = 180.0 # VL
IC2Volume = 4.0 # IC2
BindingBufferVolume = 600.0 # VEB
B_BeadsVolume = 20.0 # B-Beads
VEW1Volume = 600.0 # VEW1
VEW2Volume = 600.0 # VEW2
EtOH80pVolume = 600.0
ProtKVolume = 20.0
cRNAVolume = 4.0
IC_MS2Volume = 20.0 # MS2
ElutionBufferVolume = 100.0 # VEL
InitLysisVol = 0.0
if self.do_extraction:
if not self.add_samples: InitLysisVol += SampleVolume
if not self.add_preMix: InitLysisVol += ProtKVolume + cRNAVolume + IC_MS2Volume
if not self.add_VL: InitLysisVol += LysisBufferVolume
# Liquid classes used for pippetting.
# Others liquidClass names are defined in "protocol_steps.py"
SampleLiqClass = "Serum Asp" # = TissueHomLiqClass # SerumLiqClass="Serum Asp preMix3"
all_samples = range(NumOfSamples)
maxTips = min (self.nTips, NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
if self.do_extraction:
par = TeMag.parallelOrder(self.nTips, all_samples)
# Define the reactives in each labware (Cuvette, eppys, etc.)
# IC2 = Rtv.Reagent("IC2 - synthetic RNA " , Reactives, pos=13, volpersample= IC2Volume ,defLiqClass=W_liquidClass)
if self.add_preMix:
ProtK = Rtv.Reagent("Proteinase K ",
Reactives,
replicas=2,
pos=[15, 16], # only 16 ? pos=16
volpersample=ProtKVolume,
defLiqClass=Small_vol_disp)
cRNA = Rtv.Reagent("Carrier RNA ", Reactives, pos=14, volpersample=cRNAVolume, defLiqClass=Small_vol_disp)
IC_MS2 = Rtv.Reagent("IC MS2 phage culture ", Reactives, pos=13, volpersample=IC_MS2Volume, defLiqClass=Small_vol_disp)
pK_cRNA_MS2 = Rtv.preMix("ProtK+cRNA+IC-MS2 mix ",
Reactives,
pos=8,
components=[cRNA, ProtK, IC_MS2],
defLiqClass=W_liquidClass,
excess=20)
if self.add_VL:
LysisBuffer = Rtv.Reagent("VL - Lysis Buffer ", LysBuf, volpersample=LysisBufferVolume, defLiqClass='MN VL')
if self.do_extraction:
B_Beads = Rtv.Reagent("B - Beads ",
Reactives,
pos = [1,2],
initial_vol = 1200,
volpersample = B_BeadsVolume,
defLiqClass = Beads_LC_2,
maxFull = 70)
BindingBuffer = Rtv.Reagent("VEB - Binding Buffer ", BindBuf, volpersample=BindingBufferVolume, defLiqClass=B_liquidClass)
VEW1 = Rtv.Reagent("VEW1 - Wash Buffer ",
wt.getLabware(Lab.Trough_100ml, "4-VEW1 Wash Buffe"),
volpersample = VEW1Volume,
defLiqClass = B_liquidClass)
VEW2 = Rtv.Reagent("VEW2 - WashBuffer ",
wt.getLabware(Lab.Trough_100ml, "5-VEW2-WashBuffer" ),
volpersample =VEW2Volume,
defLiqClass =B_liquidClass)
EtOH80p = Rtv.Reagent("Ethanol 80% ",
Lab.getLabware(Lab.Trough_100ml, "7-EtOH80p" ),
volpersample=EtOH80pVolume, defLiqClass=B_liquidClass)
ElutionBuffer = Rtv.Reagent("Elution Buffer ",
ElutBuf,
volpersample =ElutionBufferVolume,
defLiqClass =B_liquidClass) # defLiqClass="Eluat" ??
# Show the CheckList GUI to the user for possible small changes
self.CheckList()
self.set_EvoMode()
# Define the reactives not shown in the CheckList GUI
# Define samples and the place for temporal reactions
for s in all_samples:
Rtv.Reagent("lysis_{:02d}".format(s + 1), Lysis, initial_vol=InitLysisVol,
pos=s + 1, defLiqClass=SampleLiqClass, excess=0)
if self.do_extraction:
Rtv.Reagent("RNA_{:02d}".format(s + 1), Eluat, initial_vol= 0.0,
pos=s+1, defLiqClass=def_liquidClass, excess=0)
if self.add_samples:
Rtv.Reagent("probe_{:02d}".format(s + 1), Samples, single_use=SampleVolume,
pos=s+1, defLiqClass=SampleLiqClass, excess=0)
Itr.wash_tips(wasteVol=30, FastWash=True).exec()
if self.do_extraction:
Te_MagS_ActivateHeater(50).exec()
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Dispense).exec()
if self.add_preMix: # add ProtK+cRNA+MS2 mix
with self.tips(tipsMask=maxMask, reuse=True, drop=False):
self.makePreMix(pK_cRNA_MS2)
self.spread ( reactive=pK_cRNA_MS2, to_labware_region= Lysis.selectOnly(all_samples))
if self.add_samples: # add samples
with self.tips(reuse=True, drop=True, preserve=True):
self.transfer( from_labware_region = Samples,
to_labware_region = Lysis,
volume = SampleVolume,
using_liquid_class = (SampleLiqClass, "Serum Disp postMix3"),
optimizeFrom = False,
optimizeTo = True,
NumSamples = NumOfSamples)
Itr.wash_tips(wasteVol=4, FastWash=True).exec()
if self.add_VL: # add LysisBuffer
with self.tips(reuse=True, drop=False): # better reuse=True, drop=False ??
self.spread ( reactive=LysisBuffer, to_labware_region= Lysis.selectOnly(all_samples))
if not self.do_extraction:
self.done()
return
with incubation(10): pass
with group("Beads binding"):
with self.tips(tipsMask=maxMask, reuse=True, drop=False):
for p in [40, 50, 60, 65]:
self.mix_reactive(B_Beads, LiqClass=Beads_LC_1, cycles=1, maxTips=maxTips, v_perc=p)
with self.tips(reuse=True, drop=True):
self.spread( reactive=B_Beads, to_labware_region=TeMag.selectOnly(all_samples))
with self.tips(reuse=True, drop=False, preserve=True, usePreserved=True):
self.wash_in_TeMag(reactive=BindingBuffer, wells=all_samples)
with self.tips(reuse=True, drop=False, preserve=True):
self.wash_in_TeMag(reactive=VEW1, wells=all_samples)
self.wash_in_TeMag(reactive=VEW2, wells=all_samples)
with self.group("Wash in TeMag with " + EtOH80p.name), self.tips():
self.spread( reactive=EtOH80p, to_labware_region= TeMag.selectOnly(all_samples))
with parallel_execution_of(mix_mag_sub, repeat=NumOfSamples//self.nTips + 1):
self.mix( TeMag.selectOnly(all_samples), EtOH80p.defLiqClass)
with incubation(minutes=0.5):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Aspirate, z_pos=24).exec()
with self.tips(usePreserved=self.preserveingTips()):
self.waste( from_labware_region= TeMag.selectOnly(all_samples))
with incubation(minutes=4):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Incubation).exec()
with incubation(minutes=4):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Aspirate, z_pos=24).exec()
self.spread( reactive=ElutionBuffer, to_labware_region=TeMag.selectOnly(all_samples))
with incubation(minutes=2):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Incubation).exec()
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Dispense).exec()
with parallel_execution_of(mix_mag_eluat, repeat=NumOfSamples//self.nTips+1):
self.mix(TeMag.selectOnly(all_samples), ElutionBuffer.defLiqClass)
with self.tips(usePreserved=self.preserveingTips(), preserve=False, drop=True):
with incubation(minutes=1.0, timer=2):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Aspirate).exec()
self.transfer( from_labware_region= TeMag.selectOnly(all_samples),
to_labware_region= Eluat.selectOnly(all_samples),
volume= ElutionBufferVolume,
optimizeTo= False,
using_liquid_class=(ElutionBuffer.defLiqClass, ElutionBuffer.defLiqClass))
self.done()
def spread(self, volume =None,
reactive =None,
to_labware_region =None,
optimize =True,
NumSamples =None,
using_liquid_class=None,
TIP_MASK =None,
num_tips =None):
"""
:param NumSamples: Priorized !!!! If true reset the selection
:param reactive: Reagent to spread
:param to_labware_region: Labware in which the destine well are selected
:param volume: if not, volume is set from the default of the source reactive
:param optimize: minimize zigzag of multi pipetting
"""
assert isinstance(reactive, Rtv.Reagent), 'A Reagent expected in reactive to spread'
assert isinstance(to_labware_region, Lab.Labware), 'A Labware expected in to_labware_region to spread'
if num_tips is None:
num_tips = self.robot.curArm().nTips # the number of tips to be used in each cycle of pipetting = all
if NumSamples:
to_labware_region.selectOnly(range(NumSamples))
else:
if not to_labware_region.selected():
to_labware_region.selectOnly(range(self.NumOfSamples))
to = to_labware_region.selected() # list of offset of selected wells
if optimize: to = to_labware_region.parallelOrder(num_tips, to)
NumSamples = len(to)
SampleCnt = NumSamples
volume = volume or reactive.volpersample
Asp_liquidClass, Dst_liquidClass = (reactive.defLiqClass, reactive.defLiqClass) if using_liquid_class is None else \
(using_liquid_class[0] or reactive.defLiqClass, using_liquid_class[1] or reactive.defLiqClass)
lf = reactive.labware
lt = to_labware_region
msg = "Spread: {v:.1f} µL of {n:s}".format(v=volume, n=reactive.name)
with group(msg):
msg += " ({v:.1f} µL total) from [grid:{fg:d} site:{fs:d} {fw:s} into {to:s}[grid:{tg:d} site:{ts:d}] in order {do:s}:" \
.format( v = reactive.minVol(),
fg = lf.location.grid,
fs = lf.location.site+1,
fw = str([str(well) for well in reactive.Replicas]) ,
do = str([i+1 for i in to]),
to = lt.label,
tg = lt.location.grid,
ts = lt.location.site+1)
Itr.comment(msg).exec()
availableDisp = 0
while SampleCnt:
if num_tips > SampleCnt: num_tips = SampleCnt
with self.tips(Rbt.tipsMask[num_tips], usePreserved=False, preserve=False): # OK want to use preserved ?? selected=??
maxMultiDisp_N = self.robot.curArm().Tips[0].type.maxVol // volume # assume all tips equal
dsp, rst = divmod(SampleCnt, num_tips)
if dsp >= maxMultiDisp_N:
dsp = maxMultiDisp_N
vol = [volume * dsp] * num_tips
availableDisp = dsp
else:
vol = [volume * (dsp + 1)] * rst + [volume * dsp] * (num_tips - rst)
availableDisp = dsp + bool(rst)
self.aspiremultiTips(num_tips, reactive, vol, LiqClass=Asp_liquidClass)
while availableDisp:
if num_tips > SampleCnt: num_tips = SampleCnt
curSample = NumSamples - SampleCnt
sel = to[curSample: curSample + num_tips] # todo what if volume > maxVol_tip ?
self.dispensemultiwells(num_tips, Dst_liquidClass, to_labware_region.selectOnly(sel), [volume] * num_tips)
availableDisp -= 1
SampleCnt -= num_tips
def makePreMix(self, preMix: Rtv.preMix,
NumSamples: int = None,
force_replies: bool = False):
"""
A preMix is just that: a premix of reactive (aka - components)
which have been already defined to add some vol per sample.
Uses one new tip per component.
It find and check self the min and max number of replica of the resulting preMix
:param preMix: what to make, predefined preMix
:param NumSamples:
:param force_replies: use all the preMix predefined replicas
:return:
"""
assert isinstance(preMix, Rtv.preMix)
robot = self.robot
mxnTips = robot.curArm().nTips # max number of Tips
ncomp = len(preMix.components)
nt = min(mxnTips, ncomp)
NumSamples = NumSamples or self.NumOfSamples
labw = preMix.labware
tVol = preMix.minVol(NumSamples)
mxnrepl = len(preMix.Replicas) # max number of replies
mnnrepl = preMix.min_num_of_replica(NumSamples) # min number of replies
assert mxnrepl >= mnnrepl, 'Please choose at least {:d} replies for {:s}'.format(mnnrepl, preMix.name)
nrepl = mxnrepl if force_replies else mnnrepl
if nrepl < mxnrepl:
print("WARNING !!! The last {:d} replies of {:s} will not be used.".format(mxnrepl-nrepl, preMix.name))
preMix.Replicas = preMix.Replicas[:nrepl]
msg = "preMix: {:.1f} µL of {:s}".format(tVol, preMix.name)
with group(msg):
msg += " into grid:{:d} site:{:d} {:s} from {:d} components:"\
.format( labw.location.grid,
labw.location.site + 1,
str([str(well) for well in preMix.Replicas]) ,
ncomp )
Itr.comment(msg).exec()
samples_per_replicas = [(NumSamples + nrepl - (ridx+1))//nrepl for ridx in range(nrepl)]
with self.tips(Rbt.tipsMask[nt]): # want to use preserved ?? selected=??
tip = -1
ctips = nt
for ridx, react in enumerate(preMix.components): # iterate reactive components
labw = react.labware
sVol = react.volpersample*preMix.excess # vol we need for each sample
rVol = sVol*NumSamples # the total vol we need of this react component
msg = " {idx:d}- {v:.1f} µL from grid:{g:d} site:{st:d}:{w:s}"\
.format( idx = ridx + 1,
v = rVol,
g = labw.location.grid,
st = labw.location.site + 1,
w = str([str(well) for well in react.Replicas]) )
Itr.comment(msg).exec()
tip += 1 # use the next tip
if tip >= nt:
ctips = min(nt, ncomp - ridx) # how many tips to use for the next gruop
tipsType = robot.curArm().Tips[0].type # only the 0 ??
self.dropTips(Rbt.tipsMask[ctips])
self.getTips(Rbt.tipsMask[ctips], tipsType)
tip = 0
mV = robot.curArm().Tips[tip].type.maxVol
# aspire/dispense multiple times if rVol don't fit in the tip (mV)
# but also if there is not sufficient reactive in the current component replica
current_comp_repl = 0
while rVol > 0:
while (react.Replicas[current_comp_repl].vol < 1): # todo define sinevoll min vol
current_comp_repl +=1
dV = min (rVol, mV, react.Replicas[current_comp_repl].vol)
self.aspire(tip, react, dV, offset=react.Replicas[current_comp_repl].offset)
self.multidispense_in_replicas(ridx, preMix, [sp/NumSamples * dV for sp in samples_per_replicas])
rVol -= dV
self.mix_reactive(preMix, maxTips=ctips)
def mix(in_labware_region,
using_liquid_class=None,
volume=None,
optimize=True):
"""
:param in_labware_region:
:param using_liquid_class:
:param volume:
:param optimize:
:return:
"""
mix_p = 0.9
in_labware_region = in_labware_region or Lab.WashWaste
assert isinstance(
in_labware_region,
Lab.Labware), 'A Labware expected in in_labware_region to be mixed'
if not volume or volume < 0.0: volume = 0.0
assert isinstance(volume, (int, float))
oriSel = in_labware_region.selected()
nt = Rbt.Robot.current.curArm(
).nTips # the number of tips to be used in each cycle of pippeting
if not oriSel:
oriSel = range(Rtv.NumOfSamples)
if optimize:
oriSel = in_labware_region.parallelOrder(nt, oriSel)
NumSamples = len(oriSel)
SampleCnt = NumSamples
if nt > SampleCnt:
nt = SampleCnt
# mV = Rbt.Robot.current.curArm().Tips[0].type.maxVol * 0.8
mV = Lab.def_DiTi.maxVol * mix_p # What tip tp use !
if volume:
v = volume
else:
v = in_labware_region.Wells[oriSel[0]].vol
v = v * mix_p
v = v if v < mV else mV
lf = in_labware_region
mx = Itr.mix(Rbt.tipsMask[nt], using_liquid_class, volume,
in_labware_region)
msg = "Mix: {v:.1f} µL of {n:s}".format(v=v, n=lf.label)
with group(msg):
msg += " [grid:{fg:d} site:{fs:d}] in order:".format(fg=lf.location.grid, fs=lf.location.site+1) \
+ str([i+1 for i in oriSel])
Itr.comment(msg).exec()
while SampleCnt:
curSample = NumSamples - SampleCnt
if nt > SampleCnt:
nt = SampleCnt
mx.tipMask = Rbt.tipsMask[nt]
sel = oriSel[curSample:curSample + nt]
spl = range(curSample, curSample + nt)
with tips(Rbt.tipsMask[nt], selected_samples=spl):
mV = Rbt.Robot.current.curArm().Tips[0].type.maxVol * mix_p
mx.labware.selectOnly(sel)
if not using_liquid_class:
if sel:
mx.liquidClass = mx.labware.selected_wells(
)[0].reactive.defLiqClass
if volume:
r = volume # r: Waste_available yet; volume: to be Waste
else:
vols = [w.vol for w in mx.labware.selected_wells()]
r_min, r_max = min(vols), max(vols)
assert r_min == r_max
r = r_max
r = r * mix_p
r = r if r < mV else mV
mx.volume = r
mx.exec()
SampleCnt -= nt
mx.labware.selectOnly(oriSel)
return oriSel
def waste(from_labware_region=None,
using_liquid_class=None,
volume=None,
to_waste_labware=None,
optimize=True):
"""
:param from_labware_region:
:param using_liquid_class:
:param volume:
:param to_waste_labware:
:param optimize:
:return:
"""
to_waste_labware = to_waste_labware or Lab.def_WashWaste
assert isinstance(
from_labware_region,
Lab.Labware), 'A Labware expected in from_labware_region to transfer'
if not volume or volume < 0.0: volume = 0.0
assert isinstance(volume, (int, float))
oriSel = from_labware_region.selected()
nt = Rbt.Robot.current.curArm(
).nTips # the number of tips to be used in each cycle of pippeting
if not oriSel:
oriSel = range(Rtv.NumOfSamples)
if optimize:
oriSel = from_labware_region.parallelOrder(nt, oriSel)
NumSamples = len(oriSel)
SampleCnt = NumSamples
if nt > SampleCnt:
nt = SampleCnt
tm = Rbt.tipsMask[nt]
nt = to_waste_labware.autoselect(maxTips=nt)
mV = Lab.def_DiTi.maxVol # todo revise !! What tip tp use !
Rest = 50 # the volume we cannot more aspire with liquid detection, to small, collisions
RestPlus = 50
CtrVol = 0.5
if volume:
v = volume
else:
v = from_labware_region.Wells[oriSel[0]].vol
Asp = Itr.aspirate(tm, Te_Mag_LC, volume, from_labware_region)
# Asp = Itr.aspirate(tm, using_liquid_class[0], volume, from_labware_region)
Dst = Itr.dispense(tm, using_liquid_class, volume, to_waste_labware)
# Ctr = Itr.moveLiha(Itr.moveLiha.y_move, Itr.moveLiha.z_start, 3.0, 2.0, tm, from_labware_region)
lf = from_labware_region
msg = "Waste: {v:.1f} µL of {n:s}".format(v=v, n=lf.label)
with group(msg):
msg += " [grid:{fg:d} site:{fs:d}] in order:".format(fg=lf.location.grid, fs=lf.location.site+1) \
+ str([i+1 for i in oriSel])
Itr.comment(msg).exec()
while SampleCnt:
curSample = NumSamples - SampleCnt
if nt > SampleCnt:
nt = SampleCnt
tm = Rbt.tipsMask[nt]
Asp.tipMask = tm
Dst.tipMask = tm
sel = oriSel[curSample:curSample + nt]
spl = range(curSample, curSample + nt)
Asp.labware.selectOnly(sel)
if volume:
r = volume # r: Waste_available yet; volume: to be Waste
else:
vols = [w.vol for w in Asp.labware.selected_wells()]
r_min, r_max = min(vols), max(vols)
assert r_min == r_max
r = r_max
if not using_liquid_class:
if sel:
Dst.liquidClass = Asp.labware.selected_wells(
)[0].reactive.defLiqClass
with tips(tm, drop=True, preserve=False, selected_samples=spl):
while r > Rest: # dont aspire Rest with these Liq Class (Liq Detect)
dV = r if r < mV else mV
if dV < Rest: break # ??
dV -= Rest # the last Rest uL have to be aspired with the other Liq Class
Asp.volume = dV # with Liq Class with Detect: ">> AVR-Serum 1000 << 365"
Dst.volume = dV
Asp.liquidClass = Te_Mag_LC # ">> AVR-Serum 1000 << 365" # "No Liq Detect"
Asp.exec()
Asp.volume = CtrVol
Asp.liquidClass = Te_Mag_Centre
with tips(allow_air=CtrVol):
Asp.exec()
if dV + Rest + RestPlus + 2 * CtrVol > mV:
Dst.exec()
r -= dV
Dst.volume = 0
else:
break
Asp.volume = Rest
Asp.liquidClass = Te_Mag_Rest # ">> AVR-Serum 1000 << 367" # "No Liq Detect"
with tips(allow_air=Rest):
Asp.exec()
Asp.volume = CtrVol
Asp.liquidClass = Te_Mag_Force_Centre
with tips(allow_air=CtrVol):
Asp.exec()
Asp.volume = RestPlus
Asp.liquidClass = Te_Mag_RestPlus # ">> AVR-Serum 1000 << 369" # "No Liq Detect"
with tips(allow_air=RestPlus):
Asp.exec()
#Ctr.exec()
Asp.volume = CtrVol
Asp.liquidClass = Te_Mag_Force_Centre
Dst.volume += Rest + RestPlus
with tips(allow_air=CtrVol):
Asp.exec()
with tips(allow_air=Rest + RestPlus):
Dst.exec()
SampleCnt -= nt
Asp.labware.selectOnly(oriSel)
Itr.wash_tips(wasteVol=4).exec()
return oriSel
def transfer(from_labware_region,
to_labware_region,
volume,
using_liquid_class=None,
optimizeFrom=True,
optimizeTo=True,
NumSamples=None):
"""
:param NumSamples: Priorized !!!! If true reset the selection
:param from_reactive: Reactive to spread
:param to_labware_region: Labware in which the destine well are selected
:param volume: if not, volume is set from the default of the source reactive
:param optimize: minimize zigzag of multipippeting
"""
assert isinstance(
from_labware_region,
Lab.Labware), 'A Labware expected in from_labware_region to transfer'
assert isinstance(
to_labware_region,
Lab.Labware), 'A Labware expected in to_labware_region to transfer'
# assert isinstance(using_liquid_class, tuple)
nt = Rbt.Robot.current.curArm(
).nTips # the number of tips to be used in each cycle of pippeting
if NumSamples: # OK? select convenient def
oriSel = range(NumSamples)
dstSel = range(NumSamples)
else:
oriSel = to_labware_region.selected()
dstSel = from_labware_region.selected()
if not dstSel:
if not oriSel:
oriSel = range(Rtv.NumOfSamples)
dstSel = range(Rtv.NumOfSamples)
else:
dstSel = oriSel
else:
if not oriSel:
oriSel = dstSel
else:
l = min(
len(oriSel), len(dstSel)
) # todo transfer the minimun of the selected ???? Best reise error
oriSel = oriSel[:l]
dstSel = dstSel[:l]
if optimizeFrom: oriSel = from_labware_region.parallelOrder(nt, oriSel)
if optimizeTo: dstSel = to_labware_region.parallelOrder(nt, dstSel)
NumSamples = len(dstSel)
SampleCnt = NumSamples
assert isinstance(volume, (int, float))
if nt > SampleCnt: nt = SampleCnt
lf = from_labware_region
lt = to_labware_region
Asp = Itr.aspirate(Rbt.tipsMask[nt],
volume=volume,
labware=from_labware_region)
Dst = Itr.dispense(Rbt.tipsMask[nt],
volume=volume,
labware=to_labware_region)
msg = "Transfer: {v:.1f} µL of {n:s}".format(v=volume, n=lf.label)
with group(msg):
msg += " [grid:{fg:d} site:{fs:d}] in order {oo:s} into {to:s}[grid:{tg:d} site:{ts:d}] in order {do:s}:" \
.format(fg=lf.location.grid, fs=lf.location.site+1, oo=str([i+1 for i in oriSel]),
do=str([i+1 for i in dstSel]), to=lt.label, tg=lt.location.grid, ts=lt.location.site+1)
Itr.comment(msg).exec()
while SampleCnt:
curSample = NumSamples - SampleCnt
if nt > SampleCnt:
nt = SampleCnt
Asp.tipMask = Rbt.tipsMask[nt]
Dst.tipMask = Rbt.tipsMask[nt]
src = oriSel[curSample:curSample + nt]
trg = dstSel[curSample:curSample + nt]
spl = range(curSample, curSample + nt)
sw = Asp.labware.selected_wells()
if isinstance(using_liquid_class, tuple):
if using_liquid_class[0]:
Asp.liquidClass = using_liquid_class[0]
else:
Asp.liquidClass = sw[0].reactive.defLiqClass
if using_liquid_class[1]:
Dst.liquidClass = using_liquid_class[1]
else:
Dst.liquidClass = sw[0].reactive.defLiqClass
else:
Asp.liquidClass = sw[0].reactive.defLiqClass
Dst.liquidClass = sw[0].reactive.defLiqClass
with tips(Rbt.tipsMask[nt], selected_samples=spl
): # todo what if volume > maxVol_tip ?
Asp.labware.selectOnly(src)
Asp.exec()
Dst.labware.selectOnly(trg)
Dst.exec()
for s, d in zip(Asp.labware.selected_wells(),
Dst.labware.selected_wells()):
d.track = s.track
SampleCnt -= nt
Asp.labware.selectOnly(oriSel)
Dst.labware.selectOnly(dstSel)
return oriSel, dstSel
def spread(volume=None,
reactive=None,
to_labware_region=None,
optimize=True,
NumSamples=None):
"""
:param NumSamples: Priorized !!!! If true reset the selection
:param reactive: Reactive to spread
:param to_labware_region: Labware in which the destine well are selected
:param volume: if not, volume is set from the default of the source reactive
:param optimize: minimize zigzag of multipippeting
"""
assert isinstance(
reactive, Rtv.Reactive), 'A Reactive expected in reactive to spread'
assert isinstance(
to_labware_region,
Lab.Labware), 'A Labware expected in to_labware_region to spread'
nt = Rbt.Robot.current.curArm(
).nTips # the number of tips to be used in each cycle of pippeting
if NumSamples:
to_labware_region.selectOnly(range(NumSamples))
else:
if not to_labware_region.selected():
to_labware_region.selectOnly(range(Rtv.NumOfSamples))
to = to_labware_region.selected()
if optimize: to = to_labware_region.parallelOrder(nt, to)
NumSamples = len(to)
SampleCnt = NumSamples
volume = volume or reactive.volpersample
if nt > SampleCnt: nt = SampleCnt
lf = reactive.labware
lt = to_labware_region
msg = "Spread: {v:.1f} µL of {n:s}".format(v=volume, n=reactive.name)
with group(msg):
msg += " {v:.1f} µL total from [grid:{fg:d} site:{fs:d} well:{fw:d}] into {to:s}[grid:{tg:d} site:{ts:d}] in order {do:s}:" \
.format(v=reactive.minVol(), fg=lf.location.grid, fs=lf.location.site+1, fw=reactive.pos+1, do=str([i+1 for i in to]),
to=lt.label, tg=lt.location.grid, ts=lt.location.site+1)
Itr.comment(msg).exec()
availableDisp = 0
while SampleCnt:
if nt > SampleCnt: nt = SampleCnt
with tips(
Rbt.tipsMask[nt], usePreserved=False,
preserve=False): # OK want to use preserved ?? selected=??
maxMultiDisp_N = Rbt.Robot.current.curArm(
).Tips[0].type.maxVol // volume # assume all tips equal
dsp, rst = divmod(SampleCnt, nt)
if dsp >= maxMultiDisp_N:
dsp = maxMultiDisp_N
vol = [volume * dsp] * nt
availableDisp = dsp
else:
vol = [volume *
(dsp + 1)] * rst + [volume * dsp] * (nt - rst)
availableDisp = dsp + bool(rst)
aspiremultiTips(nt, reactive, vol)
while availableDisp:
if nt > SampleCnt: nt = SampleCnt
curSample = NumSamples - SampleCnt
sel = to[curSample:curSample +
nt] # todo what if volume > maxVol_tip ?
dispensemultiwells(nt, reactive.defLiqClass,
to_labware_region.selectOnly(sel),
[volume] * nt)
availableDisp -= 1
SampleCnt -= nt
def Run(self):
self.set_EvoMode()
self.initialize() # set_defaults ??
wt = self.worktable
Itr.comment('Extracting RNA from {:s} samples with the MN-Vet kit'.format(str(NumOfSamples))).exec()
# Get Labwares (Cuvette, eppys, etc.) from the work table
DiTi1000_1 = wt.getLabware(Lab.DiTi_1000ul, "1000-1")
DiTi1000_2 = wt.getLabware(Lab.DiTi_1000ul, "1000-2")
DiTi1000_3 = wt.getLabware(Lab.DiTi_1000ul, "1000-3")
Reactives = wt.getLabware(Lab.GreinRack16_2mL,"Reactives" )
# Set the initial position of the tips
self.go_first_pos()
# Set volumen / sample
SampleVolume = 200.0
EtOH80pVolume = 600.0
SampleLiqClass = "Serum Asp" # = TissueHomLiqClass # SerumLiqClass="Serum Asp preMix3"
all_samples = range(NumOfSamples)
maxTips = min (self.nTips, NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
# Define the reactives in each labware (Cuvette, eppys, etc.)
VEB = Rtv.Reagent("VEB - Binding Buffer ",
BindBuf, volpersample=BindingBufferVolume, defLiqClass=B_liquidClass)
EtOH80p = Rtv.Reagent("Ethanol 80% ",
wt.getLabware(Lab.Trough_100ml, "7-EtOH80p" ),
volpersample=EtOH80pVolume, defLiqClass=B_liquidClass)
# Show the CheckList GUI to the user for possible small changes
self.CheckList()
self.set_EvoMode()
Itr.wash_tips(wasteVol=30, FastWash=True).exec()
Plate_lysis = wt.getLabware(Lab.MP96deepwell, "Plate lysis" ) # Plate 12 x 8 ?
Plate_EtOH = wt.getLabware(Lab.MP96deepwell, "Plate EtOH" ) # Plate 12 x 8 ? MP96well !!
Samples = wt.getLabware(Lab.EppRack6x16, "Proben" ) # 6x16 = 12 x 8 ?
# Define samples and the place for temporal reactions
par = Plate_lysis.parallelOrder(self.nTips, all_samples)
for s in all_samples:
Rtv.Reagent("probe_{:02d}".format(s + 1), Samples, single_use=SampleVolume,
pos=s + 1, defLiqClass=SampleLiqClass, excess=0)
Rtv.Reagent("lysis_{:02d}".format(s + 1), Plate_lysis, initial_vol=0.0, pos=par[s] + 1,
excess=0) # todo revise order !!!
Rtv.Reagent("EtOH80p_{:02d}".format(s + 1), Plate_EtOH, initial_vol=0.0, pos=par[s] + 1,
excess=0) # todo revise order !!!
with group("Prefill plate with EtOH80p"):
with tips(reuse=True, drop=False, drop_last=True):
spread(reactive=EtOH80p, to_labware_region=Plate_EtOH.selectOnly(all_samples))
dropTips()
self.done()
PanFla_119 = PrimerMix("PanFLA", ID="119", components=[Primer.IDs[320], Primer.IDs[321]])
WNV_INNT_37 = PrimerMix("WNV_5p", ID="37", components=[Primer.IDs[20], Primer.IDs[21], Primer.IDs[22]])
Exp_300 = PCRexperiment(300, "PanFLA mosq Grecia")
samples = ["Pool-1",
"Pool-2",
"Pool-3",
"Pool-4"]
Exp_300.addReactions(PanFla_119, samples)
Exp_300.addReactions(WNV_INNT_37, samples)
Exp_300.pippete_mix() # Evo75
Exp_300.pippete_samples() # Evo100
def transfer(self, from_labware_region, to_labware_region, volume, using_liquid_class=None,
optimizeFrom=True, optimizeTo=True, NumSamples=None):
"""
:param NumSamples: Priorized !!!! If true reset the selection
:param from_reactive: Reagent to spread
:param to_labware_region: Labware in which the destine well are selected
:param volume: if not, volume is set from the default of the source reactive
:param optimize: minimize zigzag of multipippeting
"""
assert isinstance(from_labware_region, Lab.Labware), 'A Labware expected in from_labware_region to transfer'
assert isinstance(to_labware_region, Lab.Labware), 'A Labware expected in to_labware_region to transfer'
# assert isinstance(using_liquid_class, tuple)
nt = self.robot.curArm().nTips # the number of tips to be used in each cycle of pippeting
if NumSamples: # OK? select convenient def
oriSel = range(NumSamples)
dstSel = range(NumSamples)
else:
oriSel = to_labware_region.selected()
dstSel = from_labware_region.selected()
if not dstSel:
if not oriSel:
oriSel = range(Rtv.NumOfSamples)
dstSel = range(Rtv.NumOfSamples)
else:
dstSel = oriSel
else:
if not oriSel:
oriSel = dstSel
else:
l = min(len(oriSel), len(dstSel)) # todo transfer the minimun of the selected ???? Best reise error
oriSel = oriSel[:l]
dstSel = dstSel[:l]
if optimizeFrom: oriSel = from_labware_region.parallelOrder(nt, oriSel)
if optimizeTo: dstSel = to_labware_region.parallelOrder(nt, dstSel)
NumSamples = len(dstSel)
SampleCnt = NumSamples
assert isinstance(volume, (int, float))
if nt > SampleCnt: nt = SampleCnt
lf = from_labware_region
lt = to_labware_region
Asp = Itr.aspirate(Rbt.tipsMask[nt], volume=volume, labware=from_labware_region)
Dst = Itr.dispense(Rbt.tipsMask[nt], volume=volume, labware=to_labware_region)
msg = "Transfer: {v:.1f} µL of {n:s}".format(v=volume, n=lf.label)
with group(msg):
msg += " [grid:{fg:d} site:{fs:d}] in order {oo:s} into {to:s}[grid:{tg:d} site:{ts:d}] in order {do:s}:" \
.format(fg =lf.location.grid,
fs =lf.location.site+1,
oo =str([i+1 for i in oriSel]),
do =str([i+1 for i in dstSel]),
to =lt.label,
tg =lt.location.grid,
ts =lt.location.site+1)
Itr.comment(msg).exec()
while SampleCnt:
curSample = NumSamples - SampleCnt
if nt > SampleCnt:
nt = SampleCnt
Asp.tipMask = Rbt.tipsMask[nt]
Dst.tipMask = Rbt.tipsMask[nt]
src = oriSel[curSample:curSample + nt]
trg = dstSel[curSample:curSample + nt]
spl = range(curSample, curSample + nt)
sw = Asp.labware.selected_wells()
if isinstance(using_liquid_class, tuple):
if using_liquid_class[0]:
Asp.liquidClass = using_liquid_class[0]
else:
Asp.liquidClass = sw[0].reactive.defLiqClass
if using_liquid_class[1]:
Dst.liquidClass = using_liquid_class[1]
else:
Dst.liquidClass = sw[0].reactive.defLiqClass
else:
Asp.liquidClass = sw[0].reactive.defLiqClass
Dst.liquidClass = sw[0].reactive.defLiqClass
with self.tips(Rbt.tipsMask[nt], selected_samples=spl): # todo what if volume > maxVol_tip ?
Asp.labware.selectOnly(src)
Asp.exec()
Dst.labware.selectOnly(trg)
Dst.exec()
for s, d in zip(Asp.labware.selected_wells(), Dst.labware.selected_wells()):
d.track = s.track
SampleCnt -= nt
Asp.labware.selectOnly(oriSel)
Dst.labware.selectOnly(dstSel)
return oriSel, dstSel
def Run(self):
self.set_EvoMode()
self.initialize() # set_defaults ??
Rtv.NumOfSamples = self.NumOfSamples
NumOfSamples = self.NumOfSamples
wt = self.worktable
Itr.comment('Extracting RNA from {:s} samples with the MN-Vet kit'.format(str(NumOfSamples))).exec()
# Get Labwares (Cuvette, eppys, etc.) from the work table
ElutBuf = wt.getLabware(Lab.Trough_100ml, "1-VEL-ElutionBuffer" )
LysBuf = wt.getLabware(Lab.Trough_100ml, "2-Vl Lysis Buffer" )
BindBuf = wt.getLabware(Lab.Trough_100ml, "3-VEB Binding Buffer" )
DiTi1000_1 = wt.getLabware(Lab.DiTi_1000ul, "1000-1")
DiTi1000_2 = wt.getLabware(Lab.DiTi_1000ul, "1000-2")
DiTi1000_3 = wt.getLabware(Lab.DiTi_1000ul, "1000-3")
Reactives = wt.getLabware(Lab.GreinRack16_2mL, "Reactives" )
# Set the initial position of the tips
self.go_first_pos()
# Set volumen / sample
SampleVolume = 200.0
LysisBufferVolume = 180.0 # VL1
IC2Volume = 5.0 # ? 4
BindingBufferVolume = 600.0
B_BeadsVolume = 20.0
VEW1Volume = 600.0
VEW2Volume = 600.0
EtOH80pVolume = 600.0
ProtKVolume = 20.0
cRNAVolume = 4.0
IC_MS2Volume = 20.0
ElutionBufferVolume = 100.0
# Liquid classes used for pippetting. Others liquidClass names are defined in "protocol_steps.py"
SampleLiqClass = "Serum Asp" # = TissueHomLiqClass # SerumLiqClass="Serum Asp preMix3"
all_samples = range(NumOfSamples)
maxTips = min (self.nTips, NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
# Define the reactives in each labware (Cuvette, eppys, etc.)
LysisBuffer = Rtv.Reagent("VL - Lysis Buffer ",
LysBuf, volpersample=LysisBufferVolume, defLiqClass=B_liquidClass)
IC2 = Rtv.Reagent("IC2 - synthetic RNA ",
Reactives, pos=11, volpersample= IC2Volume, defLiqClass=W_liquidClass)
VEB = Rtv.Reagent("VEB - Binding Buffer ",
BindBuf, volpersample=BindingBufferVolume, defLiqClass=B_liquidClass)
B_Beads = Rtv.Reagent("B - Beads ", Reactives, initial_vol=1200,
pos=1, volpersample= B_BeadsVolume, replicas=2, defLiqClass=Beads_LC_2)
VEW1 = Rtv.Reagent("VEW1 - Wash Buffer ",
wt.getLabware(Lab.Trough_100ml, "4-VEW1 Wash Buffe"),
volpersample=VEW1Volume, defLiqClass=B_liquidClass)
VEW2 = Rtv.Reagent("VEW2 - WashBuffer ",
wt.getLabware(Lab.Trough_100ml, "5-VEW2-WashBuffer" ),
volpersample=VEW2Volume, defLiqClass=B_liquidClass)
EtOH80p = Rtv.Reagent("Ethanol 80% ",
wt.getLabware(Lab.Trough_100ml, "7-EtOH80p" ),
volpersample=EtOH80pVolume, defLiqClass=B_liquidClass)
ElutionBuffer = Rtv.Reagent("Elution Buffer ",
ElutBuf, volpersample=ElutionBufferVolume, defLiqClass="Eluat")
ProtK = Rtv.Reagent("Proteinase K ",
Reactives, pos=16, volpersample= ProtKVolume, defLiqClass=Small_vol_disp)
cRNA = Rtv.Reagent("Carrier RNA ",
Reactives, pos=15, volpersample= cRNAVolume, defLiqClass=Small_vol_disp)
IC_MS2 = Rtv.Reagent("IC MS2 phage culture ",
Reactives, pos=14, volpersample= IC_MS2Volume, defLiqClass=Small_vol_disp)
pK_cRNA_MS2 = Rtv.preMix ("ProtK+cRNA+IC-MS2 mix " ,
Reactives, pos=12, components=[ ProtK, cRNA, IC2 ]
, defLiqClass=W_liquidClass, replicas=2)
# Waste = Rtv.Reagent("Waste " , self.WashWaste )
# Show the CheckList GUI to the user for posible small changes
self.CheckList()
self.set_EvoMode()
Itr.wash_tips(wasteVol=30, FastWash=True).exec()
with group("Prefill plates with VEW1, VEW2, EtOH and Elution buffer"):
Itr.userPrompt("Put the plates for VEW1, Elution Buffer and VEW2"
" in the worktable defined order").exec()
Plate_VEW1 = wt.getLabware(Lab.MP96deepwell, "Plate VEW1" ) # Plate 12 x 8 ?
Plate_ElutB = wt.getLabware(Lab.MP96well, "Plate ElutB") # Plate 12 x 8 ? MP96well !!
Plate_VEW2 = wt.getLabware(Lab.MP96deepwell, "Plate VEW2" ) # Plate 12 x 8 ?
# Define samples and the place for temporal reactions
for s in all_samples:
Rtv.Reagent("VEW1_{:02d}".format(s + 1), Plate_VEW1, initial_vol=0.0, pos=s + 1,
excess=0) # todo revise order !!!
Rtv.Reagent("VEW2_{:02d}".format(s + 1), Plate_VEW2, initial_vol=0.0, pos=s + 1,
excess=0) # todo revise order !!!
Rtv.Reagent("ElutB_{:02d}".format(s + 1), Plate_ElutB, initial_vol=0.0, pos=s + 1,
excess=0) # todo revise order !!!
with tips(reuse=True, drop=False):
spread(reactive=VEW1, to_labware_region=Plate_VEW1.selectOnly(all_samples))
with tips(reuse=True, drop=False):
spread(reactive=VEW2, to_labware_region=Plate_VEW2.selectOnly(all_samples))
with tips(reuse=True, drop=False):
spread(reactive=ElutionBuffer, to_labware_region=Plate_ElutB.selectOnly(all_samples))
Itr.userPrompt("Retire the plates for VEW1, VEW2 and Elution Buffer, and "
"Put the plates for the lysis -in place of VEW1- and for EtOH80p -in place of VEW2-").exec()
Plate_EtOH = wt.replaceWithNew(Plate_VEW2, "Plate_EtOH") # Plate 12 x 8 ?
for s in all_samples:
Rtv.Reagent("EtOH_{:02d}".format(s + 1), Plate_EtOH, initial_vol=0.0, pos=s + 1,
excess=0) # todo revise order !!!
with tips(reuse=True, drop=False):
spread(reactive=EtOH80p, to_labware_region=Plate_EtOH.selectOnly(all_samples))
with group("Sample Lysis"):
Samples = wt.getLabware(Lab.EppRack6x16, "Proben") # 6x16 = 12 x 8 ?
Plate_lysis = wt.replaceWithNew(Plate_VEW1, "Lysis_Plate") # Plate 12 x 8 ?
# Define samples and the place for temporal reactions
for s in all_samples:
Rtv.Reagent("probe_{:02d}".format(s + 1), Samples, single_use=SampleVolume,
pos=s + 1, defLiqClass=SampleLiqClass, excess=0)
Rtv.Reagent("lysis_{:02d}".format(s + 1), Plate_lysis, initial_vol=0.0, pos=s + 1,
excess=0) # todo revise order !!!
with tips(tipsMask=maxMask, reuse=True, drop=True):
pK_cRNA_MS2.make(NumOfSamples)
spread ( reactive=pK_cRNA_MS2, to_labware_region= Plate_lysis.selectOnly(all_samples))
spread ( reactive=LysisBuffer, to_labware_region= Plate_lysis.selectOnly(all_samples))
with tips(reuse=False, drop=True):
transfer( from_labware_region= Samples,
to_labware_region= Plate_lysis,
volume= SampleVolume,
using_liquid_class= (SampleLiqClass, "Serum Disp postMix3"),
optimizeFrom =False, # optimizeTo= True, # todo Really ??
NumSamples= NumOfSamples)
Itr.wash_tips(wasteVol=4, FastWash=True).exec()
# with tips(reuse=False, drop=True): # better reuse=True, drop=False, with normal Liquid Class ??
# spread ( reactive=LysisBuffer,
# to_labware_region= Plate_lysis.selectOnly(all_samples),
# using_liquid_class=(None,"Serum Disp postMix3"))
with incubation(minutes=15):
Itr.userPrompt("Please Schutteln the plates for lysis in pos 1").exec()
with group("Beads binding"):
with tips(tipsMask=maxMask, reuse=True, drop=False):
for p in [40, 50, 60, 65]:
mix_reactive(B_Beads, LiqClass=Beads_LC_1, cycles=1, maxTips=maxTips, v_perc=p)
with tips(reuse=True, drop=True):
spread( reactive=B_Beads, to_labware_region=Plate_lysis.selectOnly(all_samples))
spread( reactive=VEB, to_labware_region=Plate_lysis.selectOnly(all_samples))
# with tips(reuse=False, drop=True): # better reuse=True, drop=False, with normal Liquid Class ??
# spread ( reactive=VEB,
# to_labware_region= Plate_lysis.selectOnly(all_samples),
# using_liquid_class=(None,"Serum Disp postMix3"))
with incubation(minutes=5):
Itr.userPrompt("Please Schutteln the plates for lysis in pos 1").exec()
self.done()
def extractRNA_with_MN_Vet_Kit(NumOfSamples, CheckList=None):
Rtv.NumOfSamples = NumOfSamples
import protocols.RNAextractionMN_Mag.RobotInitRNAextraction as RI
Itr.comment('Extracting RNA from {:s} samples with the MN-Vet kit'.format(
str(NumOfSamples))).exec()
# DiTi1000_1.fill('C06')
# DiTi1000_2.fill('A11')
# DiTi1000_3.fill('A10')
Itr.set_DITI_Counter2(RI.DiTi1000_1, posInRack='A01').exec()
SampleVolume = 200.0
LysisBufferVolume = 180.0
IC2Volume = 4.0
BindingBufferVolume = 600.0
B_BeadsVolume = 20.0
VEW1Volume = 600.0
VEW2Volume = 600.0
EtOH80pVolume = 600.0
ProtKVolume = 20.0
cRNAVolume = 4.0
IC_MS2Volume = 20.0
ElutionBufferVolume = 100.0
SampleLiqClass = "Serum Asp" # = TissueHomLiqClass # SerumLiqClass="Serum Asp preMix3"
all_samples = range(Rtv.NumOfSamples)
maxTips = min(Rbt.nTips, Rtv.NumOfSamples)
maxMask = Rbt.tipsMask[maxTips]
par = RI.TeMag.parallelOrder(Rbt.nTips, all_samples)
LysisBuffer = Rtv.Reactive("VL - Lysis Buffer ",
RI.LysBuf,
volpersample=LysisBufferVolume,
defLiqClass=B_liquidClass)
IC2 = Rtv.Reactive("IC2 - synthetic RNA ",
RI.Reactives,
pos=11,
volpersample=IC2Volume,
defLiqClass=W_liquidClass)
BindingBuffer = Rtv.Reactive("VEB - Binding Buffer ",
RI.BindBuf,
volpersample=BindingBufferVolume,
defLiqClass=B_liquidClass)
B_Beads = Rtv.Reactive("B - Beads ",
RI.Reactives,
initial_vol=1200,
pos=1,
volpersample=B_BeadsVolume,
replicas=2,
defLiqClass=Beads_LC_2)
VEW1 = Rtv.Reactive("VEW1 - Wash Buffer ",
Lab.Cuvette(Lab.Trough_100ml,
Lab.Labware.Location(22, 4),
"4-VEW1 Wash Buffer"),
volpersample=VEW1Volume,
defLiqClass=B_liquidClass)
VEW2 = Rtv.Reactive("VEW2 - WashBuffer ",
Lab.Cuvette(Lab.Trough_100ml,
Lab.Labware.Location(22, 5),
"5-VEW2-WashBuffer"),
volpersample=VEW2Volume,
defLiqClass=B_liquidClass)
EtOH80p = Rtv.Reactive("Ethanol 80% ",
Lab.Cuvette(Lab.Trough_100ml,
Lab.Labware.Location(24, 1),
"7-Ethanol 80%"),
volpersample=EtOH80pVolume,
defLiqClass=B_liquidClass)
ElutionBuffer = Rtv.Reactive("Elution Buffer ",
RI.ElutBuf,
volpersample=ElutionBufferVolume,
defLiqClass="Eluat")
ProtK = Rtv.Reactive("Proteinase K ",
RI.Reactives,
pos=16,
volpersample=ProtKVolume,
defLiqClass=Small_vol_disp)
cRNA = Rtv.Reactive("Carrier RNA ",
RI.Reactives,
pos=15,
volpersample=cRNAVolume,
defLiqClass=Small_vol_disp)
IC_MS2 = Rtv.Reactive("IC MS2 phage culture ",
RI.Reactives,
pos=14,
volpersample=IC_MS2Volume,
defLiqClass=Small_vol_disp)
pK_cRNA_MS2 = Rtv.preMix("ProtK+cRNA+IC-MS2 mix ",
RI.Reactives,
pos=12,
components=[ProtK, cRNA, IC_MS2],
defLiqClass=W_liquidClass,
replicas=2)
Waste = Rtv.Reactive("Waste ", RI.WashWaste)
if CheckList is not None:
CheckList()
for s in all_samples:
Rtv.Reactive("probe_{:02d}".format(s + 1),
RI.Samples,
single_use=SampleVolume,
pos=s + 1,
defLiqClass=SampleLiqClass,
excess=0)
Rtv.Reactive("lysis_{:02d}".format(s + 1),
RI.TeMag,
initial_vol=0.0,
pos=par[s] + 1,
defLiqClass=def_liquidClass,
excess=0)
Rtv.Reactive("RNA_{:02d}".format(s + 1),
RI.Eluat,
initial_vol=0.0,
pos=s + 1,
defLiqClass=def_liquidClass,
excess=0)
Itr.wash_tips(wasteVol=30, FastWash=True).exec()
Te_MagS_ActivateHeater(50).exec()
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Dispense).exec()
with tips(tipsMask=maxMask, reuse=True, drop=False):
pK_cRNA_MS2.make()
spread(reactive=pK_cRNA_MS2,
to_labware_region=RI.TeMag.selectOnly(all_samples))
with tips(reuse=True, drop=True, preserve=True):
transfer(from_labware_region=RI.Samples,
to_labware_region=RI.TeMag,
volume=SampleVolume,
using_liquid_class=(SampleLiqClass, "Serum Disp postMix3"),
optimizeFrom=False,
optimizeTo=True,
NumSamples=Rtv.NumOfSamples)
Itr.wash_tips(wasteVol=4, FastWash=True).exec()
with tips(reuse=False, drop=True):
spread(reactive=LysisBuffer,
to_labware_region=RI.TeMag.selectOnly(all_samples))
with incubation(10):
pass
with tips(tipsMask=maxMask, reuse=True, drop=False):
for p in [40, 50, 60, 65]:
mix_reactive(B_Beads,
LiqClass=Beads_LC_1,
cycles=1,
maxTips=maxTips,
v_perc=p)
# mix_reactive(B_Beads, LiqClass=Beads_LC_2, cycles=3, maxTips=maxTips, v_perc=90)
with tips(reuse=True, drop=True):
spread(reactive=B_Beads,
to_labware_region=RI.TeMag.selectOnly(all_samples))
with tips(reuse=True, drop=False, preserve=True, usePreserved=True):
wash_in_TeMag(reactive=BindingBuffer, wells=all_samples)
with tips(reuse=True, drop=False, preserve=True):
wash_in_TeMag(reactive=VEW1, wells=all_samples)
wash_in_TeMag(reactive=VEW2, wells=all_samples)
with group("Wash in TeMag with " + EtOH80p.name), tips():
spread(reactive=EtOH80p,
to_labware_region=RI.TeMag.selectOnly(all_samples))
with parallel_execution_of(mix_mag_sub,
repeat=Rtv.NumOfSamples // Rbt.nTips +
1):
mix(RI.TeMag.selectOnly(all_samples), EtOH80p.defLiqClass)
with incubation(minutes=0.5):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Aspirate,
z_pos=24).exec()
with tips(usePreserved=preserveingTips()):
waste(from_labware_region=RI.TeMag.selectOnly(all_samples))
with incubation(minutes=4):
Te_MagS_MoveToPosition(
Te_MagS_MoveToPosition.Incubation).exec()
with incubation(minutes=4):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Aspirate,
z_pos=24).exec()
spread(reactive=ElutionBuffer,
to_labware_region=RI.TeMag.selectOnly(all_samples))
with incubation(minutes=2):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Incubation).exec()
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Dispense).exec()
with parallel_execution_of(mix_mag_eluat,
repeat=Rtv.NumOfSamples // Rbt.nTips + 1):
mix(RI.TeMag.selectOnly(all_samples), ElutionBuffer.defLiqClass)
with tips(usePreserved=preserveingTips(), preserve=False, drop=True):
with incubation(minutes=1.0, timer=2):
Te_MagS_MoveToPosition(Te_MagS_MoveToPosition.Aspirate).exec()
transfer(from_labware_region=RI.TeMag.selectOnly(all_samples),
to_labware_region=RI.Eluat.selectOnly(all_samples),
volume=ElutionBufferVolume,
optimizeTo=False,
using_liquid_class=(ElutionBuffer.defLiqClass,
ElutionBuffer.defLiqClass))