Repository 'bigwig_outlier_bed'
hg clone https://toolshed.g2.bx.psu.edu/repos/iuc/bigwig_outlier_bed

Changeset 0:ebcd48f183b3 (2024-07-05)
Next changeset 1:8377a6abb4da (2024-07-21)
Commit message:
planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/bigwig_outlier_bed commit 091caba3c5b066b293745ccee5cd31132fec3b4b
added:
README.md
bigwig_outlier_bed.py
bigwig_outlier_bed.xml
test-data/1.bigwig
test-data/bedouthi2_sample
test-data/bedouthi_sample
test-data/bedouthilo2_sample
test-data/bedouthilo_sample
test-data/bedoutlo2_sample
test-data/bedoutlo_sample
test-data/bigwig_sample
test-data/table2_sample
test-data/table_sample
b
diff -r 000000000000 -r ebcd48f183b3 README.md
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/README.md Fri Jul 05 06:00:15 2024 +0000
[
@@ -0,0 +1,31 @@
+## bigwig peak bed maker
+
+### July 30 2024 for the VGP
+
+This code will soon become a Galaxy tool, for building some of the [NIH MARBL T2T assembly polishing](https://github.com/marbl/training) tools as Galaxy workflows.
+
+JBrowse2 2.12.3 update will include a plugin for optional colours to distinguish bed features, shown being tested in the screenshots below.
+
+### Find and mark BigWig peaks to a bed file for display
+
+In the spirit of DeepTools, but finding contiguous regions where the bigwig value is either above or below a given centile.
+0.99 and 0.01 for example. These quantile cut point values are found and applied over each chromosome using some [cunning numpy code](http://gregoryzynda.com/python/numpy/contiguous/interval/2019/11/29/contiguous-regions.html)
+
+![image](https://github.com/fubar2/bigwig_peak_bed/assets/6016266/cdee3a2b-ae31-4282-b744-992c15fb49db)
+
+![image](https://github.com/fubar2/bigwig_peak_bed/assets/6016266/59d1564b-0c34-42a3-b437-44332cf1b2f0)
+
+Big differences between chromosomes 14,15,21,22 and Y in this "all contigs" view - explanations welcomed:
+
+![image](https://github.com/fubar2/bigwig_peak_bed/assets/6016266/162bf681-2977-4eb8-8d6f-9dad5b3931f8)
+
+
+[pybedtools](https://github.com/jackh726/bigtools) is used for the bigwig interface. Optionally allow
+multiple bigwigs to be processed into a single bed - the bed features have the bigwig name in the label for viewing.
+
+### Note on quantiles per chromosome rather than quantiles for the whole bigwig
+
+It is just not feasible to hold all contigs in the entire decoded bigwig in RAM to estimate quantiles. It may be
+better to sample across all chromosomes so as not to lose any systematic differences between them - the current method will hide those
+differences unfortunately. Sampling might be possible. Looking at the actual quantile values across a couple of test bigwigs suggests that
+there is not much variation between chromosomes but there's now a tabular report to check them for each input bigwig.
b
diff -r 000000000000 -r ebcd48f183b3 bigwig_outlier_bed.py
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/bigwig_outlier_bed.py Fri Jul 05 06:00:15 2024 +0000
[
@@ -0,0 +1,177 @@
+"""
+Ross Lazarus June 2024 for VGP
+Bigwigs are great, but hard to reliably "see" small low coverage or small very high coverage regions.
+Colouring in JB2 tracks will need a new plugin, so this code will find bigwig regions above and below a chosen percentile point.
+0.99 and 0.01 work well in testing with a minimum span of 10 bp.
+Multiple bigwigs **with the same reference** can be combined - bed segments will be named appropriately
+Combining multiple references works but is silly because only display will rely on one reference so others will not be shown...
+Tricksy numpy method from http://gregoryzynda.com/python/numpy/contiguous/interval/2019/11/29/contiguous-regions.html
+takes about 95 seconds for a 17MB test wiggle
+JBrowse2 bed normally displays ignore the score, so could provide separate low/high bed file outputs as an option.
+Update june 30 2024: wrote a 'no-build' plugin for beds to display red/blue if >0/<0 so those are used for scores
+Bed interval naming must be short for JB2 but needs input bigwig name and (lo or hi).
+"""
+
+import argparse
+import copy
+import os
+import sys
+from pathlib import Path
+
+import numpy as np
+import pybigtools
+
+
+class findOut:
+
+    def __init__(self, args):
+        self.bwnames = args.bigwig
+        self.bwlabels = args.bigwiglabels
+        self.bedwin = args.minwin
+        self.qlo = args.qlo
+        self.qhi = args.qhi
+        self.outbeds = args.outbeds
+        self.bedouthi = args.bedouthi
+        self.bedoutlo = args.bedoutlo
+        self.bedouthilo = args.bedouthilo
+        self.tableoutfile = args.tableoutfile
+        self.bedwin = args.minwin
+        self.qhi = args.qhi
+        self.qlo = args.qlo
+        nbw = len(args.bigwig)
+        nlab = len(args.bigwiglabels)
+        if nlab < nbw:
+            self.bwlabels += ["Nolabel"] * (nbw - nlab)
+        self.makeBed()
+
+    def processVals(self, bw, isTop):
+        """
+        idea from http://gregoryzynda.com/python/numpy/contiguous/interval/2019/11/29/contiguous-regions.html
+        Fast segmentation into regions by taking np.diff on the boolean array of over (under) cutpoint indicators in bwex.
+        This only gives non-zero values at the segment boundaries where there's a change, so those zeros are all removed in bwexdnz
+        leaving an array of segment start/end positions. That's twisted around into an array of start/end coordinates.
+        Magical. Fast. Could do the same for means or medians over windows for sparse bigwigs like repeat regions.
+        """
+        if isTop:
+            bwex = np.r_[False, bw >= self.bwtop, False]  # extend with 0s
+        else:
+            bwex = np.r_[False, bw <= self.bwbot, False]
+        bwexd = np.diff(bwex)
+        bwexdnz = bwexd.nonzero()[0]
+        bwregions = np.reshape(bwexdnz, (-1, 2))
+        return bwregions
+
+    def writeBed(self, bed, bedfname):
+        """
+        potentially multiple
+        """
+        bed.sort()
+        beds = ["%s\t%d\t%d\t%s\t%d" % x for x in bed]
+        with open(bedfname, "w") as bedf:
+            bedf.write("\n".join(beds))
+            bedf.write("\n")
+
+    def makeTableRow(self, bw, bwlabel, chr):
+        """
+        called for every contig, but messy inline
+        """
+        bwmean = np.mean(bw)
+        bwstd = np.std(bw)
+        bwmax = np.max(bw)
+        nrow = np.size(bw)
+        bwmin = np.min(bw)
+        row = "%s\t%s\t%d\t%f\t%f\t%f\t%f" % (
+            bwlabel,
+            chr,
+            nrow,
+            bwmean,
+            bwstd,
+            bwmin,
+            bwmax,
+        )
+        if self.qhi is not None:
+            row += "\t%f" % self.bwtop
+        else:
+            row += "\t"
+        if self.qlo is not None:
+            row += "\t%f" % self.bwbot
+        else:
+            row += "\t"
+        return row
+
+    def makeBed(self):
+        bedhi = []
+        bedlo = []
+        bwlabels = self.bwlabels
+        bwnames = self.bwnames
+        if self.tableoutfile:
+            restab = ["bigwig\tcontig\tn\tmean\tstd\tmin\tmax\tqtop\tqbot"]
+        for i, bwname in enumerate(bwnames):
+            bwlabel = bwlabels[i].replace(" ", "")
+            fakepath = "in%d.bw" % i
+            if os.path.isfile(fakepath):
+                os.remove(fakepath)
+            p = Path(fakepath)
+            p.symlink_to(bwname)  # required by pybigtools (!)
+            bwf = pybigtools.open(fakepath)
+            chrlist = bwf.chroms()
+            chrs = list(chrlist.keys())
+            chrs.sort()
+            for chr in chrs:
+                bw = bwf.values(chr)
+                bw = bw[~np.isnan(bw)]  # some have NaN if parts of a contig not covered
+                if self.qhi is not None:
+                    self.bwtop = np.quantile(bw, self.qhi)
+                    bwhi = self.processVals(bw, isTop=True)
+                    for j, seg in enumerate(bwhi):
+                        if seg[1] - seg[0] >= self.bedwin:
+                            bedhi.append((chr, seg[0], seg[1], "%s_hi" % (bwlabel), 1))
+                if self.qlo is not None:
+                    self.bwbot = np.quantile(bw, self.qlo)
+                    bwlo = self.processVals(bw, isTop=False)
+                    for j, seg in enumerate(bwlo):
+                        if seg[1] - seg[0] >= self.bedwin:
+                            bedlo.append((chr, seg[0], seg[1], "%s_lo" % (bwlabel), -1))
+                if self.tableoutfile:
+                    row = self.makeTableRow(bw, bwlabel, chr)
+                    restab.append(copy.copy(row))
+        if self.tableoutfile:
+            stable = "\n".join(restab)
+            with open(self.tableoutfile, "w") as t:
+                t.write(stable)
+                t.write("\n")
+        some = False
+        if self.qlo:
+            if self.outbeds in ["outall", "outlo", "outlohi"]:
+                self.writeBed(bedlo, self.bedoutlo)
+                some = True
+        if self.qhi:
+            if self.outbeds in ["outall", "outlohi", "outhi"]:
+                self.writeBed(bedhi, self.bedouthi)
+                some = True
+        if self.outbeds in ["outall", "outhilo"]:
+            allbed = bedlo + bedhi
+            self.writeBed(allbed, self.bedouthilo)
+            some = True
+        if not some:
+            sys.stderr.write(
+                "Invalid configuration - no output could be created. Was qlo missing and only low output requested for example?"
+            )
+            sys.exit(2)
+
+
+if __name__ == "__main__":
+    parser = argparse.ArgumentParser()
+    a = parser.add_argument
+    a("-m", "--minwin", default=10, type=int)
+    a("-l", "--qlo", default=None, type=float)
+    a("-i", "--qhi", default=None, type=float)
+    a("--bedouthi", default=None)
+    a("--bedoutlo", default=None)
+    a("--bedouthilo", default=None)
+    a("-w", "--bigwig", nargs="+")
+    a("-n", "--bigwiglabels", nargs="+")
+    a("-o", "--outbeds", default="outhilo", help="optional high and low combined bed")
+    a("-t", "--tableoutfile", default=None)
+    args = parser.parse_args()
+    findOut(args)
b
diff -r 000000000000 -r ebcd48f183b3 bigwig_outlier_bed.xml
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/bigwig_outlier_bed.xml Fri Jul 05 06:00:15 2024 +0000
[
b'@@ -0,0 +1,177 @@\n+<tool name="Bigwig extremes to bed features" id="bigwig_outlier_bed" version="0.1.4" profile="22.05">\n+  <description>Writes high and low bigwig runs as features in a bed file</description>\n+  <edam_topics>\n+      <edam_topic>topic_0157</edam_topic>\n+      <edam_topic>topic_0092</edam_topic>\n+  </edam_topics>\n+  <edam_operations>\n+      <edam_operation>operation_0337</edam_operation>\n+  </edam_operations>\n+  <xrefs>\n+    <xref type="bio.tools">bigtools</xref>\n+  </xrefs>\n+  <requirements>\n+    <requirement type="package" version="3.12.3">python</requirement>\n+    <requirement type="package" version="2.0.0">numpy</requirement>\n+    <requirement type="package" version="0.1.4">pybigtools</requirement>\n+  </requirements>\n+  <required_files>\n+      <include path="bigwig_outlier_bed.py"/>\n+  </required_files>\n+  <version_command><![CDATA[python -c "import pybigtools; from importlib.metadata import version; print(version(\'pybigtools\'))"]]></version_command>\n+  <command><![CDATA[python \'${__tool_directory__}/bigwig_outlier_bed.py\'\n+--bigwig \n+#for bw in $bigwig: \n+ \'$bw\' \n+#end for\n+--bigwiglabels\n+#for bw in $bigwig: \n+  \'$bw.name\'\n+#end for\n+--outbeds \'$outbeds\'\n+#if $outbeds in [\'outhilo\', \'outall\']:\n+  --bedouthilo \'$bedouthilo\'\n+#end if\n+#if $outbeds in [\'outhi\', \'outall\', \'outlohi\']:\n+  --bedouthi \'$bedouthi\'\n+#end if\n+#if $outbeds in [\'outlo\', \'outall\', \'outlohi\']:\n+  --bedoutlo \'$bedoutlo\'\n+#end if\n+--minwin \'$minwin\'\n+#if $qhi:\n+--qhi \'$qhi\'\n+#end if\n+#if $qlo:\n+--qlo \'$qlo\'\n+#end if\n+#if $tableout == "create":\n+ --tableoutfile \'$tableoutfile\'\n+#end if\n+]]></command>\n+  <inputs>\n+    <param name="bigwig" type="data" optional="false" label="Choose one or more bigwig file(s) to return outlier regions as a bed file" \n+      help="If more than one, MUST all use the same reference sequence to be displayable. Feature names will include the bigwig label." format="bigwig" multiple="true"/>\n+    <param name="minwin" type="integer" value="10" label="Minimum continuous bases to count as a high or low bed feature" \n+      help="Continuous features as long or longer than this window size will appear as bed features"/>\n+    <param name="qhi" type="float" value="0.99" label="Quantile cutoff for a high region - 0.99 will cut off at or above the 99th percentile" help="Required" optional="false"/>\n+    <param name="qlo" type="float" value="0.01" label="Quantile cutoff for a low region - 0.01 will cut off at or below the 1st percentile." help="Optional" optional="true"/>\n+    <param name="outbeds" type="select" label="Select the required bed file outputs" help="Any combination of the 3 different kinds of bed file output can be made">\n+      <option value="outhilo" selected="true">Make 1 bed output with both low and high regions</option>\n+      <option value="outhi">Make 1 bed output with high regions only</option>\n+      <option value="outlo">Make 1 bed output with low regions only</option>\n+      <option value="outall">Make 3 bed outputs with low and high together in one, high in one and low in the other</option>\n+      <option value="outlohi">Make 2 bed outputs with high in one and low in the other</option>\n+    </param>\n+    <param name="tableout" type="select" label="Write a table showing contig statistics for each bigwig input" help="">\n+      <option value="donotmake">Do not create this report</option>\n+      <option value="create" selected="true">Create this report</option>\n+    </param>\n+  </inputs>\n+  <outputs>\n+    <data name="bedouthilo" format="bed" label="High_and_low_bed" hidden="false">\n+      <filter>outbeds in ["outall", "outhilo"]</filter>\n+    </data>\n+    <data name="bedouthi" format="bed" label="High bed" hidden="false">\n+      <filter>outbeds in ["outall", "outlohi", "outhi"]</filter>\n+    </data>\n+    <data name="bedoutlo" format="bed" label="Low bed" hidden="false">\n+      <filter>outbeds in ["outall", "outlohi", "outlo"]</filter>\n+    </data>\n+    <data name="tableoutfile" format="tabular" label="Con'..b'pect_num_outputs="3">\n+      <output name="bedouthi" value="bedouthi_sample" compare="diff" lines_diff="0"/>\n+      <output name="bedoutlo" value="bedoutlo_sample" compare="diff" lines_diff="0"/>\n+      <output name="tableoutfile" value="table_sample" compare="diff" lines_diff="0"/>\n+      <param name="outbeds" value="outlohi"/>\n+      <param name="bigwig" value="bigwig_sample"/>\n+      <param name="minwin" value="10"/>\n+      <param name="qhi" value="0.99"/>\n+      <param name="qlo" value="0.01"/>\n+      <param name="tableout" value="create"/>\n+    </test>\n+    <test expect_num_outputs="4">\n+      <output name="bedouthilo" value="bedouthilo2_sample" compare="diff" lines_diff="0"/>\n+      <output name="bedoutlo" value="bedoutlo2_sample" compare="diff" lines_diff="0"/>\n+      <output name="bedouthi" value="bedouthi2_sample" compare="diff" lines_diff="0"/>\n+      <output name="tableoutfile" value="table2_sample" compare="diff" lines_diff="0"/>\n+      <param name="outbeds" value="outall"/>\n+      <param name="bigwig" value="bigwig_sample,1.bigwig"/>\n+      <param name="minwin" value="10"/>\n+      <param name="qhi" value="0.99"/>\n+      <param name="qlo" value="0.01"/>\n+      <param name="tableout" value="create"/>\n+    </test>\n+  </tests>\n+  <help><![CDATA[\n+\n+ **Purpose**\n+ \n+ *Combine bigwig outlier regions into bed files*\n+ \n+ Bigwigs allow quantative tracks to be viewed in an interactive genome browser like JBrowse2. \n+ Peaks are easy to see. Unusually low regions can be harder to spot, even if they are relatively large, unless the view is zoomed right in.\n+ Automated methods for combining evidence from multiple bigwigs can be useful for constructing browseable *issues* or other kinds of summary bed format tracks.\n+ For example, combining coverage outlier regions, with the frequency of specific dicnucleotide short tandem repeats,\n+ for evaluating technical sequencing technology effects in the evaluation of a genome assembly described at https://github.com/arangrhie/T2T-Polish\n+\n+ **What does it produce?**\n+\n+ Bed format results are output, containing each continuous segment of at least *minwin* base pairs above a cut point, or below another cut point. \n+ These can be viewed as features on the reference genome using a genome browser tool like JBrowse2.\n+ Three kinds of bed files can be created depending on the values included.  \n+ Both high and low regions in one bed output is the default. This can be displayed in JBrowse2 with colour indicating the high or low status, \n+ one less track and a little easier to understand. High and low features can be output as separate bed files.\n+\n+ **How is it controlled?**\n+\n+ The cut points are calculated using a user supplied quantile, from each chromosome\'s bigwig value distribution. \n+ The defaults are 0.99 and 0.01 and the default *minwin* is 10.  \n+ The probability of 10 values at or below the 1st percentile purely by chance is about 0.01**10, so false positives should be\n+ rare, even in a 3GB genome.  \n+ This data driven and non-parametric method is preferred for the asymmetrical distributions found in typical bigwigs, such as depth of coverage \n+ for genome sequencing reads. Coverage values are truncated at zero, and regions with very high values often form a long sparse right tail. \n+\n+ **How do I choose the input data?**\n+\n+ One or more bigwigs and can be selected as inputs.  \n+ Multiple bigwigs will be combined in bed files, so must share the reference genome to display\n+ using JBrowse2.\n+ \n+ .. class:: warningmark\n+\n+ **Lower quantile may not behave as expected in bigwigs with large fractions of zero values**\n+\n+ The lower cut point may be problematic for integer values like coverage if many values are zero. For example, if 5% of bases have zero coverage, the 1st percentile is also zero, \n+ but that cut point will include the entire 5% *at or below 0*\n+\n+ \n+  ]]></help>\n+  <citations>\n+    <citation type="doi">10.1093/bioinformatics/btae350</citation>\n+  </citations>\n+</tool>\n+\n'
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diff -r 000000000000 -r ebcd48f183b3 test-data/1.bigwig
b
Binary file test-data/1.bigwig has changed
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diff -r 000000000000 -r ebcd48f183b3 test-data/bedouthi2_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
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diff -r 000000000000 -r ebcd48f183b3 test-data/bedouthi_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
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diff -r 000000000000 -r ebcd48f183b3 test-data/bedouthilo2_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
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b
diff -r 000000000000 -r ebcd48f183b3 test-data/bedouthilo_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/test-data/bedouthilo_sample Fri Jul 05 06:00:15 2024 +0000
b
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b
diff -r 000000000000 -r ebcd48f183b3 test-data/bedoutlo2_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/test-data/bedoutlo2_sample Fri Jul 05 06:00:15 2024 +0000
b
@@ -0,0 +1,91 @@
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b
diff -r 000000000000 -r ebcd48f183b3 test-data/bedoutlo_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/test-data/bedoutlo_sample Fri Jul 05 06:00:15 2024 +0000
b
@@ -0,0 +1,69 @@
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b
diff -r 000000000000 -r ebcd48f183b3 test-data/bigwig_sample
b
Binary file test-data/bigwig_sample has changed
b
diff -r 000000000000 -r ebcd48f183b3 test-data/table2_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/test-data/table2_sample Fri Jul 05 06:00:15 2024 +0000
b
@@ -0,0 +1,3 @@
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b
diff -r 000000000000 -r ebcd48f183b3 test-data/table_sample
--- /dev/null Thu Jan 01 00:00:00 1970 +0000
+++ b/test-data/table_sample Fri Jul 05 06:00:15 2024 +0000
b
@@ -0,0 +1,2 @@
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