def test_get_absolute_scale(track_data):
from fluff.util import get_absolute_scale
assert 10.0 == get_absolute_scale(10, track_data)
assert 99.0 == get_absolute_scale(99.0, track_data)
assert 99.0 == get_absolute_scale("99.0", track_data)
assert 83.5 == get_absolute_scale("90.0%", track_data, False)
assert 7.0 == get_absolute_scale("10%", track_data)
assert type([]) == type(get_absolute_scale("80%", track_data, True))
for x, y in zip([82.4, 74.8, 75.8], get_absolute_scale("80%", track_data, True)):
assert round(x - y, 5) == 0
def test_get_absolute_scale(track_data):
from fluff.util import get_absolute_scale
assert 10.0 == get_absolute_scale(10, track_data)
assert 99.0 == get_absolute_scale(99.0, track_data)
assert 99.0 == get_absolute_scale("99.0", track_data)
assert 83.5 == get_absolute_scale("90.0%", track_data, False)
assert 7.0 == get_absolute_scale("10%", track_data)
assert type([]) == type(get_absolute_scale("80%", track_data, True))
for x, y in zip([82.4, 74.8, 75.8],
get_absolute_scale("80%", track_data, True)):
assert round(x - y, 5) == 0
def test_get_absolute_scale_low(track_data_low):
from fluff.util import get_absolute_scale
assert 5.0 == get_absolute_scale("10%", track_data_low)
assert 5.0 == get_absolute_scale("50%", track_data_low)
def heatmap(args):
datafiles = args.datafiles
for x in args.datafiles:
if not os.path.isfile(x):
print("ERROR: Data file '{0}' does not exist".format(x))
sys.exit(1)
for x in args.datafiles:
if '.bam' in x and not os.path.isfile("{0}.bai".format(x)):
print(
"Data file '{0}' does not have an index file. Creating an index file for {0}."
.format(x))
pysam.index(x)
# Options Parser
featurefile = args.featurefile
datafiles = [x.strip() for x in args.datafiles]
tracks = [os.path.basename(x) for x in datafiles]
titles = [os.path.splitext(x)[0] for x in tracks]
colors = parse_colors(args.colors)
bgcolors = parse_colors(args.bgcolors)
outfile = args.outfile
extend_up = args.extend
extend_down = args.extend
fragmentsize = args.fragmentsize
cluster_type = args.clustering[0].lower()
merge_mirrored = args.merge_mirrored
bins = (extend_up + extend_down) / args.binsize
rmdup = args.rmdup
rpkm = args.rpkm
rmrepeats = args.rmrepeats
ncpus = args.cpus
distancefunction = args.distancefunction.lower()
dynam = args.graphdynamics
fontsize = args.textfontsize
colorbar = args.colorbar
seed = args.seed
# Check for mutually exclusive parameters
if dynam:
if merge_mirrored:
print("ERROR: -m and -g option CANNOT be used together")
sys.exit(1)
if distancefunction == 'e':
print(
'Dynamics can only be identified using Pearson correlation as metric.'
)
print('Assigning metric to Pearson correlation')
distancefunction = 'p'
# Method of clustering
if (args.pick != None):
pick = [i - 1 for i in split_ranges(args.pick)]
if not all(i <= len(tracks) - 1 for i in pick):
sys.stderr.write(
"You picked a non-existent file for clustering.\n")
sys.exit(1)
else:
pick = list(range(len(datafiles)))
if not cluster_type in ["k", "h", "n"]:
sys.stderr.write("Unknown clustering type!\n")
sys.exit(1)
# Number of clusters
if cluster_type == "k" and not args.numclusters >= 2:
sys.stderr.write("Please provide number of clusters!\n")
sys.exit(1)
# Distance function
if not distancefunction in ["euclidean", "pearson"]:
sys.stderr.write("Unknown distance function!\n")
sys.exit(1)
else:
METRIC = distancefunction
print("{} distance method".format(METRIC))
## Get scale for each track
tscale = [1.0 for track in datafiles]
# Function to load heatmap data
def load_data(featurefile,
amount_bins,
extend_dyn_up,
extend_dyn_down,
rmdup,
rpkm,
rmrepeats,
fragmentsize,
dynam,
guard=None):
if guard is None:
guard = []
# Calculate the profile data
data = {}
regions = []
print("Loading data")
try:
# Load data in parallel
pool = multiprocessing.Pool(processes=ncpus)
jobs = []
for datafile in datafiles:
jobs.append(
pool.apply_async(load_heatmap_data,
args=(featurefile, datafile, amount_bins,
extend_dyn_up, extend_dyn_down,
rmdup, rpkm, rmrepeats,
fragmentsize, dynam, guard)))
for job in jobs:
track, regions, profile, guard = job.get()
data[track] = profile
except Exception as e:
sys.stderr.write("Error loading data in parallel, trying serial\n")
sys.stderr.write("Error: {}\n".format(e))
for datafile in datafiles:
track, regions, profile, guard = load_heatmap_data(
featurefile, datafile, amount_bins, extend_dyn_up,
extend_dyn_down, rmdup, rpkm, rmrepeats, fragmentsize,
dynam, guard)
data[track] = profile
return data, regions, guard
# -g : Option to try and get dynamics
# Extend features 1kb up/down stream
# Cluster them in one bin
# Cluster them in one bin
guard = []
amount_bins = bins
extend_dyn_up = extend_up
extend_dyn_down = extend_down
if dynam:
# load the data once to get the features which extend below 0
guard = check_data(featurefile, extend_dyn_up, extend_dyn_down)
extend_dyn_up = 1000
extend_dyn_down = 1000
amount_bins = 1
# Load data for clustering
data, regions, guard = load_data(featurefile, amount_bins, extend_dyn_up,
extend_dyn_down, rmdup, rpkm, rmrepeats,
fragmentsize, dynam, guard)
# Normalize
norm_data = normalize_data(data, cfg.DEFAULT_PERCENTILE)
clus = hstack([
norm_data[t] for i, t in enumerate(tracks) if (not pick or i in pick)
])
ind, labels = cluster_profile(clus,
cluster_type=cluster_type,
numclusters=args.numclusters,
dist=METRIC,
random_state=seed)
if cluster_type == "k":
if not dynam and merge_mirrored:
(i, j) = mirror_clusters(data, labels)
while j:
for track in list(data.keys()):
data[track][labels == j] = [
row[::-1] for row in data[track][labels == j]
]
for k in range(len(regions)):
if labels[k] == j:
(chrom, start, end, gene, strand) = regions[k]
if strand == "+":
strand = "-"
else:
strand = "+"
regions[k] = (chrom, start, end, gene, strand)
n = len(set(labels))
labels[labels == j] = i
for k in range(j + 1, n):
labels[labels == k] = k - 1
(i, j) = mirror_clusters(data, labels)
# Load data for visualization if -g option was used
if dynam:
data, regions, guard = load_data(featurefile, bins, extend_up,
extend_down, rmdup, rpkm, rmrepeats,
fragmentsize, dynam, guard)
f = open("{0}_clusters.bed".format(outfile), "w")
for (chrom, start, end, gene, strand), cluster in zip(
array(regions, dtype="object")[ind],
array(labels)[ind]):
if not gene:
f.write("{0}\t{1}\t{2}\t.\t{3}\t{4}\n".format(
chrom, start, end, cluster + 1, strand))
else:
f.write("{0}\t{1}\t{2}\t{3}\t{4}\t{5}\n".format(
chrom, start, end, gene, cluster + 1, strand))
f.close()
# Save read counts
readcounts = {}
for i, track in enumerate(tracks):
readcounts[track] = {}
readcounts[track]['bins'] = []
for idx, row in enumerate(data[track]):
bins = ''
for b in row:
if not bins:
bins = '{0}'.format(b)
else:
bins = '{0};{1}'.format(bins, b)
readcounts[track]['bins'].append(bins)
input_fileBins = open('{0}_readCounts.txt'.format(outfile), 'w')
input_fileBins.write('Regions\t')
for i, track in enumerate(titles):
input_fileBins.write('{0}\t'.format(track))
input_fileBins.write('\n')
for i, track in enumerate(tracks):
for idx in ind:
input_fileBins.write('{0}:{1}-{2}\t'.format(
regions[idx][0], regions[idx][1], regions[idx][2]))
for i, track in enumerate(tracks):
input_fileBins.write('{0}\t'.format(
readcounts[track]['bins'][idx]))
input_fileBins.write('\n')
break
input_fileBins.close()
if not cluster_type == "k":
labels = None
scale = get_absolute_scale(args.scale, [data[track] for track in tracks])
heatmap_plot(data, ind[::-1], outfile, tracks, titles, colors, bgcolors,
scale, tscale, labels, fontsize, colorbar)
def heatmap(args):
datafiles = args.datafiles
for x in args.datafiles:
if not os.path.isfile(x):
print("ERROR: Data file '{0}' does not exist".format(x))
sys.exit(1)
for x in args.datafiles:
if '.bam' in x and not os.path.isfile("{0}.bai".format(x)):
print("Data file '{0}' does not have an index file. Creating an index file for {0}.".format(x))
pysam.index(x)
# Options Parser
featurefile = args.featurefile
datafiles = [x.strip() for x in args.datafiles]
tracks = [os.path.basename(x) for x in datafiles]
titles = [os.path.splitext(x)[0] for x in tracks]
colors = parse_colors(args.colors)
bgcolors = parse_colors(args.bgcolors)
outfile = args.outfile
extend_up = args.extend
extend_down = args.extend
fragmentsize = args.fragmentsize
cluster_type = args.clustering[0].lower()
merge_mirrored = args.merge_mirrored
bins = (extend_up + extend_down) / args.binsize
rmdup = args.rmdup
rpkm = args.rpkm
rmrepeats = args.rmrepeats
ncpus = args.cpus
distancefunction = args.distancefunction.lower()
dynam = args.graphdynamics
fontsize = args.textfontsize
colorbar = args.colorbar
seed = args.seed
# Check for mutually exclusive parameters
if dynam:
if merge_mirrored:
print("ERROR: -m and -g option CANNOT be used together")
sys.exit(1)
if distancefunction == 'e':
print('Dynamics can only be identified using Pearson correlation as metric.')
print('Assigning metric to Pearson correlation')
distancefunction = 'p'
# Method of clustering
if (args.pick != None):
pick = [i - 1 for i in split_ranges(args.pick)]
if not all(i <= len(tracks) - 1 for i in pick):
sys.stderr.write("You picked a non-existent file for clustering.\n")
sys.exit(1)
else:
pick = list(range(len(datafiles)))
if not cluster_type in ["k", "h", "n"]:
sys.stderr.write("Unknown clustering type!\n")
sys.exit(1)
# Number of clusters
if cluster_type == "k" and not args.numclusters >= 2:
sys.stderr.write("Please provide number of clusters!\n")
sys.exit(1)
# Distance function
if not distancefunction in ["euclidean", "pearson"]:
sys.stderr.write("Unknown distance function!\n")
sys.exit(1)
else:
METRIC = distancefunction
print("{} distance method".format(METRIC))
## Get scale for each track
tscale = [1.0 for track in datafiles]
# Function to load heatmap data
def load_data(featurefile, amount_bins, extend_dyn_up, extend_dyn_down, rmdup, rpkm, rmrepeats, fragmentsize, dynam,
guard=None):
if guard is None:
guard = []
# Calculate the profile data
data = {}
regions = []
print("Loading data")
try:
# Load data in parallel
pool = multiprocessing.Pool(processes=ncpus)
jobs = []
for datafile in datafiles:
jobs.append(pool.apply_async(load_heatmap_data, args=(
featurefile, datafile, amount_bins, extend_dyn_up, extend_dyn_down, rmdup, rpkm, rmrepeats,
fragmentsize, dynam, guard)))
for job in jobs:
track, regions, profile, guard = job.get()
data[track] = profile
except Exception as e:
sys.stderr.write("Error loading data in parallel, trying serial\n")
sys.stderr.write("Error: {}\n".format(e))
for datafile in datafiles:
track, regions, profile, guard = load_heatmap_data(featurefile, datafile, amount_bins, extend_dyn_up,
extend_dyn_down, rmdup, rpkm, rmrepeats,
fragmentsize, dynam, guard)
data[track] = profile
return data, regions, guard
# -g : Option to try and get dynamics
# Extend features 1kb up/down stream
# Cluster them in one bin
# Cluster them in one bin
guard = []
amount_bins = bins
extend_dyn_up = extend_up
extend_dyn_down = extend_down
if dynam:
# load the data once to get the features which extend below 0
guard = check_data(featurefile, extend_dyn_up, extend_dyn_down)
extend_dyn_up = 1000
extend_dyn_down = 1000
amount_bins = 1
# Load data for clustering
data, regions, guard = load_data(featurefile, amount_bins, extend_dyn_up, extend_dyn_down, rmdup, rpkm,
rmrepeats,
fragmentsize, dynam, guard)
# Normalize
norm_data = normalize_data(data, cfg.DEFAULT_PERCENTILE)
clus = hstack([norm_data[t] for i, t in enumerate(tracks) if (not pick or i in pick)])
ind, labels = cluster_profile(
clus,
cluster_type=cluster_type,
numclusters = args.numclusters,
dist=METRIC,
random_state=seed)
if cluster_type == "k":
if not dynam and merge_mirrored:
(i, j) = mirror_clusters(data, labels)
while j:
for track in list(data.keys()):
data[track][labels == j] = [row[::-1] for row in data[track][labels == j]]
for k in range(len(regions)):
if labels[k] == j:
(chrom, start, end, gene, strand) = regions[k]
if strand == "+":
strand = "-"
else:
strand = "+"
regions[k] = (chrom, start, end, gene, strand)
n = len(set(labels))
labels[labels == j] = i
for k in range(j + 1, n):
labels[labels == k] = k - 1
(i, j) = mirror_clusters(data, labels)
# Load data for visualization if -g option was used
if dynam:
data, regions, guard = load_data(featurefile, bins, extend_up, extend_down, rmdup, rpkm, rmrepeats,
fragmentsize, dynam, guard)
f = open("{0}_clusters.bed".format(outfile), "w")
for (chrom, start, end, gene, strand), cluster in zip(array(regions, dtype="object")[ind], array(labels)[ind]):
if not gene:
f.write("{0}\t{1}\t{2}\t.\t{3}\t{4}\n".format(chrom, start, end, cluster + 1, strand))
else:
f.write("{0}\t{1}\t{2}\t{3}\t{4}\t{5}\n".format(chrom, start, end, gene, cluster + 1, strand))
f.close()
# Save read counts
readcounts = {}
for i, track in enumerate(tracks):
readcounts[track] = {}
readcounts[track]['bins'] = []
for idx, row in enumerate(data[track]):
bins = ''
for b in row:
if not bins:
bins = '{0}'.format(b)
else:
bins = '{0};{1}'.format(bins, b)
readcounts[track]['bins'].append(bins)
input_fileBins = open('{0}_readCounts.txt'.format(outfile), 'w')
input_fileBins.write('Regions\t')
for i, track in enumerate(titles):
input_fileBins.write('{0}\t'.format(track))
input_fileBins.write('\n')
for i, track in enumerate(tracks):
for idx in ind:
input_fileBins.write('{0}:{1}-{2}\t'.format(regions[idx][0], regions[idx][1], regions[idx][2]))
for i, track in enumerate(tracks):
input_fileBins.write('{0}\t'.format(readcounts[track]['bins'][idx]))
input_fileBins.write('\n')
break
input_fileBins.close()
if not cluster_type == "k":
labels = None
scale = get_absolute_scale(args.scale, [data[track] for track in tracks])
heatmap_plot(data, ind[::-1], outfile, tracks, titles, colors, bgcolors, scale, tscale, labels, fontsize, colorbar)
def test_get_absolute_scale_low(track_data_low):
from fluff.util import get_absolute_scale
assert 5.0 == get_absolute_scale("10%", track_data_low)
assert 5.0 == get_absolute_scale("50%", track_data_low)