def load_crosses(self, seq_id, cross_id, field):
# Not available via intake 22 Oct 2020
crosses_path = self.release_dir / "site_filters" / "crosses_stats"
crosses_store = self.gcs.get_mapper(crosses_path.as_posix())
crosses_group = zarr.Group(crosses_store)
return da.from_zarr(crosses_group[seq_id][field][cross_id])
def _benchmark_load_zarr_datasets(self, zarr_paths):
callsets = []
self.benchmark_profiler.start_benchmark(
operation_name="Load Zarr Dataset")
for zarr_path in zarr_paths:
store = zarr.DirectoryStore(zarr_path)
callset = zarr.Group(store=store, read_only=True)
callsets.append(callset)
self.benchmark_profiler.end_benchmark()
return callsets
def open_group(path):
if path.endswith("h5"):
return h5py.File(path, "r")
elif path.endswith(("zarr2", "zarr")):
return zarr.open_group(path, "r")
elif path.endswith("zip"):
zz = zarr.ZipStore(path)
return zarr.Group(zz)
else:
raise ValueError(
"Bad filepath provided: {0}. Only hdf5/zarr supported.".format(
path))
def execute(cls, ctx, op):
import zarr
fs = get_fs(op.path, None)
fs_map = FSMap(op.path, fs)
group = zarr.Group(store=fs_map, path=op.group)
array = group[op.dataset]
to_store = ctx[op.inputs[0].key]
axis_offsets = op.axis_offsets
shape = to_store.shape
array[tuple(
slice(offset, offset + size)
for offset, size in zip(axis_offsets, shape))] = to_store
ctx[op.outputs[0].key] = np.empty((0, ) * to_store.ndim,
dtype=to_store.dtype)
def load_variants_array(self, seq_id, field="POS", mask=None):
"""
release_pa
"""
path = self.release_dir / "snp_genotypes" / "all" / "sites"
# need to open as mapping if this on cloud
storez = self.gcs.get_mapper(path.as_posix())
calldata = zarr.Group(storez)
arr = da.from_zarr(calldata[f"{seq_id}/variants/{field}"])
if mask is not None:
assert isinstance(mask, da.core.Array), "mask must be a dask_array"
arr = da.compress(mask, arr, axis=0).compute_chunk_sizes()
return arr
def load_calldata_by_sampleset(self,
seq_id,
sampleset,
field="GT",
mask=None):
if isinstance(sampleset, str):
path = self.release_dir / "snp_genotypes" / "all" / sampleset
print(path)
# need to open as mapping if this on cloud
storez = self.gcs.get_mapper(path.as_posix())
calldata = zarr.Group(storez)
arr = da.from_zarr(calldata[f"{seq_id}/calldata/{field}"])
elif isinstance(sampleset, list):
arr = da.concatenate([
self.load_calldata_by_sampleset(
seq_id, s, field=field, mask=None) for s in sampleset
],
axis=1)
else:
raise ValueError(
"sampleset must be a string, or a list of strings")
if mask is not None:
assert isinstance(mask, da.core.Array), "mask must be a dask_array"
arr = da.compress(mask, arr, axis=0).compute_chunk_sizes()
if field == "GT":
arr = allel.GenotypeDaskArray(arr)
return arr
def main():
parser = argparse.ArgumentParser(
description=
'This script takes a single zarr zipstore, and estimates the contamination rate, providing a log'
'likelihood ratio vs the null model')
parser.add_argument(
'--input',
required=True,
help=
'Path to zarr file containing genotypes and allele depths, zipped Zarr file with data for a single sample.'
'This should follow the standard format of {sample}/{seqid}/calldata/GT and {sample}/{seqid}/calldata/AD.'
)
parser.add_argument(
'--sites',
required=True,
help=
'Path to zarr describing which sites in `input` were genotyped. This is used to match the `input` to the'
'allele frequencies below. variants/POS is required.')
parser.add_argument(
'--allele-frequencies',
required=True,
help=
'path to zarr file describing allele frequencies. This has two purposes: 1) to select SNPs to downsample'
'to, based on the `minimum_af` argument. 2) To provide a prior expectation on the frequency of genotypes.'
'The first level of the zarr file should be groups for seqids, with each containing `POS` (position) and'
'AF (allele frequencies). The shape of the AF array must be Nx4, where N is the size of the 1D POS array.'
'The order of alleles *must* correspond to the coding in the input data. There is no requirement to have a'
'similar shape to the input genotypes, although a minimum level of intersection is required!'
)
parser.add_argument('--seqid',
required=True,
nargs='+',
help='name of chromosome(s) or contig(s) to process. ')
parser.add_argument('--output',
required=True,
help='path to output file stem')
parser.add_argument('--downsample',
required=False,
default=20000,
help='number of sites to consider.',
type=int)
parser.add_argument(
'--minimum-af',
required=False,
default=0.05,
help=
'minimum minor allele frequency in reference population to consider. Sites with higher MAF are more '
'powerful at detecting contamination',
type=float)
parser.add_argument('--sequence-error-rate',
required=False,
default=1e-3,
help='probability of observing a non REF/ALT base',
type=float)
parser.add_argument(
'--minimum-coverage',
required=False,
default=10,
help=
'minimum read depth to use. Low depths have low power to detect contamination',
type=int)
parser.add_argument('--plot', dest='plot', action='store_true')
parser.add_argument('--no-plot', dest='plot', action='store_false')
parser.add_argument('--log', dest='log', action='store_true')
parser.add_argument('--no-log', dest='log', action='store_false')
parser.set_defaults(plot=True, log=False)
try:
args = {
"input": snakemake.input.input,
"sites": snakemake.input.sites,
"allele_frequencies": snakemake.input.allele_frequencies,
"seqid": snakemake.params.seqid,
"output": snakemake.params.stem,
"minimum_af": snakemake.params.minimum_af,
"minimum_coverage": snakemake.params.minimum_coverage,
"sequence_error_rate": snakemake.params.seq_err_rate,
"downsample": snakemake.params.downsample,
"plot": snakemake.params.plot,
"log": snakemake.params.log
}
log("Args read via snakemake")
except NameError:
args = vars(parser.parse_args())
log("Args read via command line")
seqids = args['seqid']
sequence_error_rate = args['sequence_error_rate']
downsample_n = args["downsample"]
minimum_minor_af = args["minimum_af"]
output_csv = args['output'] + ".contamination.csv"
output_png = args['output'] + ".allele_balance.png"
output_log = args["output"] + ".{alpha}.log"
sample_store = zarr.ZipStore(args["input"], mode="r")
sample_callset = zarr.Group(sample_store)
sites = zarr.ZipStore(args["sites"], mode="r")
variant_sites = zarr.Group(sites)
sample = next(iter(sample_callset))
concatenated_sample_callset, _ = concatenate_arrays(
sample_callset[sample], seqids, paths=["calldata/GT", "calldata/AD"])
gt = allel.GenotypeArray(concatenated_sample_callset["calldata/GT"])
ad = concatenated_sample_callset["calldata/AD"]
concatenated_sites, concatenated_site_shapes = concatenate_arrays(
variant_sites, seqids, ["variants/POS"])
pos = concatenated_sites["variants/POS"]
assert pos.shape[0] == gt.shape[0] == ad.shape[
0], "Shape inconsistency. {0}, {1}, {2}".format(
pos.shape, gt.shape, ad.shape)
# load allele frequencies required to compute weights
allele_frequencies_z = zarr.open_group(args['allele_frequencies'], "r")
concatenated_af_arrays, concatenated_af_shapes = concatenate_arrays(
allele_frequencies_z, seqids, ["POS", "AF"])
af_pos = concatenated_af_arrays["POS"]
# This is a 2D array of the frequency of the ALT allele in some other dataset.
af_val = concatenated_af_arrays["AF"]
assert af_val.shape[
1] == 4, "Allele frequencies must contain all 4 alleles, even if unobserved."
# for the sample_gt: Keep if
# a) in af, b) is_called and c) is_biallelic
# step 1 find the intersection this works on multi indexes
loc_gt, loc_af = locate_intersection(pos, concatenated_site_shapes, af_pos,
concatenated_af_shapes)
flt_af_val = np.compress(loc_af, af_val, axis=0)
flt_gt = np.compress(loc_gt, gt, axis=0)
flt_ad = np.compress(loc_gt, ad, axis=0)
# now we need to filter both by is biallelic and is called.
is_bial_ref_pop = np.count_nonzero(flt_af_val, axis=1) == 2
is_called = flt_gt.is_called()[:, 0]
# compress the intersection by the AND of these
keep_loc = is_called & is_bial_ref_pop
alt_frequency_pass = np.compress(keep_loc, flt_af_val, axis=0)
allele_depth_pass = np.compress(keep_loc, flt_ad, axis=0)
# recode the allele depth to 0/1.
# find the "alt" column.
log("Ordering alleles by frequency for REF/ALT/ERR")
min_cov_reached = allele_depth_pass[:, 0].sum(
axis=1) >= args['minimum_coverage']
ix_cols_sort = np.argsort(alt_frequency_pass, axis=1)[:, ::-1]
# indices of all rows
ix_rows = np.arange(alt_frequency_pass.shape[0])
# apply the sorting operation
allele_depth_pass_reordered = np.squeeze(allele_depth_pass)[
ix_rows[:, np.newaxis], ix_cols_sort]
# Define allele counts: sum final 2 columns, representing ref/alt/error
allele_depths = allele_depth_pass_reordered[:, :3]
allele_depths[:,
2] = allele_depths[:, 2] + allele_depth_pass_reordered[:, 3]
assert allele_depths.shape[1] == 3
# issue with some samples having a third allele (ie not in phase 2) discovered at high frequency
# Filter sites where more than 10% of reads look like errors.
probably_biallelic = allele_depth_pass_reordered[:, 2] < (
.1 * allele_depth_pass_reordered.sum(axis=1))
# step 2 create the 0/1/2 from the allele frequencies.
major_af = alt_frequency_pass.max(axis=1)
# select the values with the highest MAF.
log("Selecting variants on which to perform analysis")
while True:
eligible = probably_biallelic & min_cov_reached & (
(1 - major_af) > minimum_minor_af)
if eligible.sum() > downsample_n:
break
minimum_minor_af -= 0.01
if minimum_minor_af < 0:
log("Insufficient variants meet criteria to compute contamination. n={0}, min={1}"
.format(eligible.sum(), downsample_n))
break
res = pd.DataFrame(index=[sample], columns=["LLR", "LL", "pc_contam"])
if eligible.sum() > downsample_n:
log("Downsample from {0} to {1}".format(eligible.sum(), downsample_n))
ix_ds = np.sort(
np.random.choice(np.where(eligible)[0], size=downsample_n))
major_af = np.take(major_af, ix_ds, axis=0)
allele_depths = np.take(allele_depths, ix_ds, axis=0)
genotype_weights = np.log(determine_weights(major_af))
log("estimating contamination...")
xv = minimize_scalar(compute_likelihood,
args=(sequence_error_rate, allele_depths,
genotype_weights, args["log"], output_log),
bounds=(0, 0.5),
method="Bounded",
options={"xatol": 1e-6})
# compute the likelihood at alpha = 0, to report likelihood ratio.
null = compute_likelihood(0.0, sequence_error_rate, allele_depths,
genotype_weights, args["log"], output_log)
# return the llr / ll / estimate
res.loc[sample] = -min(xv.fun - null, 0), -xv.fun, xv.x * 100
if args['plot']:
plot_allele_balance(flt_gt, flt_ad, output_png, res.iloc[0])
res.to_csv(output_csv)
import gzip
import pandas as pd
import os
import gcsfs
import zarr
from itertools import compress
gcs_bucket_fs = gcsfs.GCSFileSystem(project='malariagen-jupyterhub',
token='anon',
access='read_only')
geno_bi_path = os.path.join(
"ag1000g-release/phase2.AR1/variation/main/zarr/biallelic/ag1000g.phase2.ar1.pass.biallelic"
)
gcsacmap = gcs_bucket_fs.get_mapper(root=geno_bi_path)
callset_biallel = zarr.Group(gcsacmap, read_only=True)
metadata = pd.read_csv("samples.meta.txt", sep="\t")
pop_selection = metadata.population.isin({
'GHcol', 'GHgam', 'BFgam', 'BFcol', 'GM', 'GW', 'GNgam', 'GNcol', 'CIcol'
}).values
callset_fn = callset_biallel
def get_consecutive_true(a):
if a.sum() == 0:
return 0
else:
return np.diff(
np.where(np.concatenate(
([a[0]], a[:-1] != a[1:], [True])))[0])[::2].max()
def load_mask(self, seq_id, mask_id, filters_model="dt_20200416"):
mask_path = self.release_dir / "site_filters" / filters_model / mask_id
mask_store = self.gcs.get_mapper(mask_path.as_posix())
mask_group = zarr.Group(mask_store)
return da.from_zarr(mask_group[seq_id]["variants/filter_pass"])
def open_zarr_dataset(zarr_path):
store = zarr.DirectoryStore(zarr_path)
callset = zarr.Group(store=store, read_only=True)
return callset
def load_filter_n(self, seq_id):
path = self.release_dir / "accessibility" / "accessibility.zarr"
storez = self.gcs.get_mapper(path.as_posix())
zarrdata = zarr.Group(storez)
return da.from_zarr(zarrdata[f"{seq_id}/filter_n"])
def main():
parser = argparse.ArgumentParser(
description=
'Given a zipped Zarr store summarize information about the coverage/calls'
)
parser.add_argument(
'--input',
required=True,
help='path to Zarr store containing calldata for a single sample')
parser.add_argument('--seqid',
required=False,
help='name of chromosomes or contigs to process',
nargs="+",
default=["2R", "2L", "3R", "3L", "X"],
type=str)
parser.add_argument(
'--output',
required=True,
help='path to output basename for stats table and coverage histogram')
try:
args = {
"input": snakemake.input.input,
"seqid": snakemake.params.seqid,
"output": snakemake.params.stem
}
log("Args read via snakemake")
except NameError:
args = vars(parser.parse_args())
log("Args read via command line")
zfn = args['input']
store = zarr.ZipStore(zfn, mode="r")
callset = zarr.Group(store)
csv_out = args['output'] + ".callstats.csv"
npy_out = args['output'] + ".covhist.npz"
# Holders for data
df_cols = [
"nSitesCalled", "nHomRef", "nHet", "nHetRef", "nHomAlt",
"nNonRefAlleles"
]
ad_df = pd.DataFrame(index=args['seqid'], columns=df_cols)
cov_hist = dict()
sample = next(iter(callset))
for chrom in args['seqid']:
gt = allel.GenotypeArray(callset[sample][chrom]["calldata/GT"])
is_called = np.squeeze(gt.is_called())
# if no coverage at all
if is_called.sum() == 0:
ad_df.loc[chrom] = 0
cov_hist[chrom] = np.array([is_called.shape[0]])
continue
all_allele_depths = callset[sample][chrom]["calldata/AD"][:, 0]
# To calculate DP at each position/sample we need to filter missing genotpyes.
allele_depths_with_cov = np.compress(is_called,
all_allele_depths,
axis=0)
# Histogram of coverage
sum_by_alt = allele_depths_with_cov.sum(axis=1)
bc = np.bincount(sum_by_alt, minlength=251)
bc[0] = (~is_called).sum()
cov_hist[chrom] = bc
nHomRef = gt.count_hom_ref()
nHet = gt.count_het()
nHetRef = gt.count_het(allele=0)
nHomAlt = gt.count_hom_alt()
nNonRefAlleles = (2 * nHomAlt) + nHetRef + (2 * (nHet - nHetRef))
total_calls = is_called.sum()
ad_df.loc[chrom] = [
total_calls, nHomRef, nHet, nHetRef, nHomAlt, nNonRefAlleles
]
ad_df.to_csv(csv_out)
np.savez(npy_out, **cov_hist)
