transformation_plate = p.ref("transformation_plate",
cont_type="96-pcr",
discard=True)
lb_media = "rs17bafcbmyrmh"
growth_plate = p.ref("growth_plate", cont_type="96-deep", storage="cold_4")
#culture plate layout:
#0:lb 1:amp 2:amp/ara
#3:lb 4:amp 5:amp/ara
#0:no_pglo 1:pglo 2:pglo
#3:no_pglo 4:no_pglo 5:pglo
p.transfer(transform_sln.well(0), transformation_plate.wells_from(0, 2),
"150:ul")
p.incubate(transformation_plate, "cold_4", "5:minute")
p.autopick(ecoli_source.all_wells(), transformation_plate.wells_from(0, 2))
p.transfer(pglo.well(0), transformation_plate.well(0), "10:ul")
for w in transformation_plate.wells_from(0, 2):
p.mix(w, volume="75:microliter")
sources = WellGroup([transformation_plate.well(0)] * 2 +
[transformation_plate.well(1)] * 2)
dests = WellGroup(
transformation_plate.wells_from(12, 2, columnwise=True) +
transformation_plate.wells_from(13, 2, columnwise=True))
p.transfer(sources, dests, "50:ul")
p.thermocycle(transformation_plate, [{
"cycles":
1,
"steps": [{
#well 9-12: no bactera, no ara (control)
num_picks = 4
selected_well = 2
grow_vol = 250 #ul
ara_vol = round(grow_vol / 32)
grow_plate = p.ref("grow_plate", cont_type="96-flat", storage="cold_4")
agar = p.ref("ct1amxs8eggsk7", cont_type="6-flat", storage="cold_4")
p.provision("rs18s8x4qbsvjz", grow_plate.wells_from(0, num_picks * 3),
str(grow_vol) +
":microliter") #lb liquid with amp. +2 for controls
arabinose = p.ref("arabinose", cont_type="micro-1.5", storage="ambient")
p.provision("rs185rp3d22ca5", arabinose.wells(0),
str(ara_vol * (num_picks) + 25) +
": microliter") #10% L(+)-Arabinose
#p.image_plate(agar, mode="top", dataref="agar_plate")
p.autopick(agar.well(selected_well), grow_plate.wells_from(0, num_picks * 2))
p.transfer(arabinose.wells(0), grow_plate.wells_from(0, num_picks),
str(ara_vol) + ":microliter")
p.incubate(grow_plate, "warm_37", "24:hour", shaking=True)
# platereader settings based on http://ww3.tecan.com/platform/apps/virtualdirectories/gcm/downloads/AN_INF200PRO_enhanced_FI_Bottom_V1_0.pdf
p.fluorescence(grow_plate,
grow_plate.wells_from(0, num_picks * 3),
excitation="483:nanometer",
emission="535:nanometer",
dataref="grow_plate")
print json.dumps(p.as_dict(), indent=2)