public rna seq

compute4/RNA-seq_genotype_calling/parameters.csv

@@ -32,4 +32,4 @@ SNVMix,/target/gpfs2/gcc/home/dasha/tools/SNVMix2-0.11.8-r5/SNVMix2,,,
 GenotypeCallingJar,/target/gpfs2/gcc/home/dasha/scripts/genotyping/GenotypeCalling/dist/GenotypeCalling.jar,,,
 snpList,/target/gpfs2/gcc/home/dasha/resources/hg19/genotypes/1000G/all_snp_positions.txt,,,
 shapeitversion,v2.644,,,
-shapeitBin,${tooldir}/Shapeit-${shapeitversion}/shapeit.v2.r644.linux.x86_64,,string,
+shapeitBin,${tooldir}/Shapeit-${shapeitversion}/shapeit.v2.r644.linux.x86_64,,string,

compute4/RNA-seq_genotype_calling/protocols/ConvertSNVMixToGen.ftl

@@ -48,34 +48,34 @@ do
 done
 
 
-# ${JAVA_HOME}/bin/java \
-        # -Xmx4g \
-        # -jar /target/gpfs2/gcc/home/dasha/scripts/genotyping/GenotypeCalling/dist/GenotypeCalling.jar \
-        # --mode SNVMixToGen \
-        # --fileList ${genotypeFolder}/fileList.txt \
-        # --p-value 0.8 \
-        # --out ${genotypeFolder}/___tmp___chr
-
-# returnCode=$?
-# echo "Return code ${returnCode}"
-
-# if [ "${returnCode}" -eq "0" ]
-# then
+ ${JAVA_HOME}/bin/java \
+         -Xmx4g \
+         -jar /target/gpfs2/gcc/home/dasha/scripts/genotyping/GenotypeCalling/dist/GenotypeCalling.jar \
+         --mode SNVMixToGen \
+         --fileList ${genotypeFolder}/fileList.txt \
+         --p-value 0.8 \
+         --out ${genotypeFolder}/___tmp___chr
+
+ returnCode=$?
+ echo "Return code ${returnCode}"
+
+ if [ "${returnCode}" -eq "0" ]
+ then
 
-	# echo "Moving temp files: ${genotypeFolder}/___tmp___chr* to ${genotypeFolder}/chr*"
-	# tmpFiles="${genotypeFolder}/___tmp___chr*"
-	# for f in $tmpFiles
-	# do
-		# mv $f ${f//___tmp___/}
-	# done
+	 echo "Moving temp files: ${genotypeFolder}/___tmp___chr* to ${genotypeFolder}/chr*"
+	 tmpFiles="${genotypeFolder}/___tmp___chr*"
+	 for f in $tmpFiles
+	 do
+		 mv $f ${f//___tmp___/}
+	 done
 
-# else
+ else
 
-	# echo -e "\nNon zero return code not making files final. Existing temp files are kept for debugging purposes\n\n"
-	Return non zero return code
-	# exit 1
+	 echo -e "\nNon zero return code not making files final. Existing temp files are kept for debugging purposes\n\n"
+	#Return non zero return code
+	 exit 1
 
-# fi
+ fi
 
 chrTriTyperDirs=""
 
@@ -97,18 +97,18 @@ do
 		genFileSorted=${genFile//.gen/.sorted.gen}
 
 
-		# sort -k3,3n ${genFile} > ${genFileSorted}
+		sort -k3,3n ${genFile} > ${genFileSorted}
 
 		genFileSortedFiltered=${genFile//.gen/_CR0.8_maf0.01.gen}
 
-		# /target/gpfs2/gcc/tools/qctool/qctool_v1.3-linux-x86_64/qctool \
-		# -g $genFileSorted \
-		# -s ${sampleFile} \
-		# -og ${genFileSortedFiltered} \
-		# -maf 0.01 1 \
-		# -hwe 4 \
-		# -snp-missing-rate 0.8 \
-		# -omit-chromosome 
+		/target/gpfs2/gcc/tools/qctool/qctool_v1.3-linux-x86_64/qctool \
+		-g $genFileSorted \
+		-s ${sampleFile} \
+		-og ${genFileSortedFiltered} \
+		-maf 0.01 1 \
+		-hwe 4 \
+		-snp-missing-rate 0.8 \
+		-omit-chromosome 
 
 		trityperFolder=${genFile%.gen}
 		mkdir -p ${trityperFolder}

compute4/RNA-seq_genotype_calling/protocols/CreateSNVMixFileList.ftl

@@ -0,0 +1,48 @@
+#MOLGENIS walltime=1:00:00 nodes=1 cores=1 mem=4
+
+#FOREACH mergedStudy
+
+genotypeFolder="${genotypeFolder}"
+JAVA_HOME="${JAVA_HOME}"
+
+declare -a samples=(${ssvQuoted(sample)})
+declare -a snvmixOuts=(${ssvQuoted(snvmixOut)})
+<#noparse>
+
+mkdir -p ${genotypeFolder}
+
+echo "genotypeFolder=${genotypeFolder}"
+echo "snvMixOuts=${snvmixOuts[*]}"
+echo "samples=${samples[*]}"
+
+rm -f ${genotypeFolder}/fileList.txt
+
+
+
+declare -a samplesProcessed=()
+
+for (( i = 0 ; i < ${#samples[@]} ; i++ )) 
+do
+
+	for processedSample in ${samplesProcessed[@]}
+	do
+		if [ $processedSample == ${samples[$i]} ]
+		then
+			continue 2
+		fi
+	done
+
+	samplesProcessed=("${samplesProcessed[@]}" "${samples[$i]}")
+	echo -e "sample:${samples[$i]}\tgenotype file:${snvmixOuts[$i]}"
+
+	if [ -f ${snvmixOuts[$i]} ]
+	then
+		echo -e  "${samples[$i]}\t${snvmixOuts[$i]}" >> ${genotypeFolder}/fileList.txt
+	else
+		echo "Skipping sample ${samples[$i]} no snvmix output"
+	fi
+
+
+done
+
+</#noparse>

compute4/RNA-seq_genotype_calling/protocols/SNVMix.ftl

@@ -82,7 +82,10 @@ returnCode=$?
 
 echo "return code snvMix ${returnCode}"
 
-if [ $returnCode -eq 0 ]
+
+ count=`cut -f 1 -d ":" ${snvmixOut}___tmp___ | uniq | wc -l`
+
+if [ $count -ge 22 ]
 then
 
 	echo "Moving temp file: ${snvmixOut}___tmp___ to $snvmixOut"

compute4/RNA-seq_genotype_calling/workflowCall.csv

@@ -0,0 +1,2 @@
+name,protocol_name,PreviousSteps_name
+SNVMix,SNVMix,

compute4/RNA-seq_genotype_calling/workflowCreateFileList.csv

@@ -0,0 +1,2 @@
+name,protocol_name,PreviousSteps_name
+CreateSNVMixFileList,CreateSNVMixFileList,

compute4/RNA-seq_genotype_calling/workflowMerge.csv

@@ -0,0 +1,2 @@
+name,protocol_name,PreviousSteps_name
+ConvertSNVMixToGen,ConvertSNVMixToGen,

compute4/RNA-seq_quantify_fluxCapacitor/parameters.csv

@@ -19,7 +19,7 @@ run,,,,
 baseFolder,,,,
 sortedBam,${baseFolder}/${studyEna}/${sample}/${run}/${run}Aligned.out.sorted.bam,,,
 expressionFolder,${baseFolder}/${mergedStudy}/expressionData/,,,
-gtfExpression,"${baseFolder}/${studyEna}/${sample}/${sample}.flux.gtf",,,
+gtfExpression,"${baseFolder}/${studyEna}/${sample}/${run}/${run}.flux.gtf",,,
 expressionTable,${expressionFolder}/expression_table.transcr.v71.flux.txt,,,
 #,,,,
 annotationGtf,/target/gpfs2/gcc/home/dasha/resources/hg19/v71/Homo_sapiens.GRCh37.71.cut.sorted.gtf,,,

compute4/RNA-seq_quantify_fluxCapacitor/protocols/FluxCapacitor.ftl

@@ -1,4 +1,4 @@
-#MOLGENIS walltime=24:00:00 nodes=1 cores=1 mem=6
+#MOLGENIS walltime=24:00:00 nodes=1 cores=2 mem=7
 
 bamToBed="${bamToBed}"
 sortedBam="${sortedBam}"

compute4/RNA-seq_quantify_fluxCapacitor/protocols/MakeExpressionTable.ftl

@@ -1,4 +1,4 @@
-#MOLGENIS walltime=6:00:00 nodes=1 cores=1 mem=4
+#MOLGENIS walltime=24:00:00 nodes=1 cores=1 mem=4
 
 #FOREACH mergedStudy
 mkdir -p ${expressionFolder}

compute4/RNA-seq_quantify_gene_level/parameters.csv

@@ -1,16 +1,16 @@
 Name,defaultValue,description,dataType,hasOne_name
 clusterQueue,gcc,,,
 scheduler,PBS,,,
-mem,4,Memory in GB,,
-walltime,6:00:00,,,
+mem,6,Memory in GB,,
+walltime,24:00:00,,,
 cores,1,,,
 defaultInterpreter,#!/bin/bash,,,
 jobname,jobname,,string,
 #,,,,
 home,/target/gpfs2/gcc/home/dasha/,,,
 root,/target/gpfs2/gcc/,the root to your tools and data,string,
-bashrc,${root}/gcc.bashrc,,,
-toolDir,${root}tools/,root Dir for tools,string,
+bashrc,/gcc/groups/gcc/home/gcc.bashrc,,,
+toolDir,/gcc/tools/,root Dir for tools,string,
 #,,,,
 studyEna,,,,
 mergedStudy,,,,expressionFolder
@@ -19,8 +19,8 @@ run,,,,
 baseFolder,,,,
 sortedBam,${baseFolder}/${studyEna}/${sample}/${run}/${run}Aligned.out.sorted.bam,,,
 expressionFolder,${baseFolder}/${mergedStudy}/expressionData/,,,
-txtExpression,"${baseFolder}/${studyEna}/${sample}/${sample}.htseq.txt",,,
-expressionTable,${expressionFolder}/expression_table.transcr.v71.htseq.txt,,,
+txtExpression,"${baseFolder}/${studyEna}/${sample}/${run}/${run}.htseq.txt",,,
+expressionTable,${expressionFolder}/expression_table.genelevel.v71.htseq.txt,,,
 #,,,,
 annotationGtf,/target/gpfs2/gcc/home/dasha/resources/hg19/v71/Homo_sapiens.GRCh37.71.cut.sorted.gtf,,,
 geneAnnotationTxt,/target/gpfs2/gcc/home/dasha/resources/hg19/annotations/v71/annotation_geneIds_v71.txt.gz,,,
@@ -30,3 +30,4 @@ JAVA_HOME,${toolDir}/jdk/,,,
 samtools,${toolDir}samtools-0.1.18/samtools,,,
 htseq_count,/target/gpfs2/gcc/home/dasha/tools/HTSeq-0.5.4p3/HTSeq/scripts/count.py,,,
 processReadCountsJar,/target/gpfs2/gcc/home/dasha/scripts/processReadCounts/ProcessReadCounts/dist/ProcessReadCounts.jar,,,
+python,python,,,

compute4/RNA-seq_quantify_gene_level/protocols/HTSeq_count.ftl

@@ -1,37 +1,48 @@
-#MOLGENIS walltime=20:00:00 nodes=1 cores=1 mem=6
+#MOLGENIS walltime=24:00:00 nodes=1 cores=1 mem=6
 
 sortedBam="${sortedBam}"
 htseq_count="${htseq_count}"
 annotationGtf="${annotationGtf}"
 txtExpression="${txtExpression}"
 samtools=${samtools}
+python=${python}
 
 <#noparse>
 
 echo -e "sortedBam=${sortedBam}\nannotationGtf=${annotationGtf}\ntxtExpression=${txtExpression}"
 
+alloutputsexist ${txtExpression}
 
 echo "Sorting bam file by name"
 
-${samtools} sort \
--n \
-${sortedBam} \
-${sortedBam%bam}byName
+${samtools} \
+	sort \
+	-n \
+	${sortedBam} \
+	${TMPDIR}/nameSorted.bam
 
 
 echo -e "\nQuantifying expression"
 
-${samtools} \
+if ${samtools} \
 	view -h \
-	${sortedBam%bam}byName.bam | \
-/target/gpfs2/gcc/tools/Python-2.7.3/bin/python \
-${htseq_count} \
+	${TMPDIR}/nameSorted.bam | \
+	/target/gpfs2/gcc/tools/Python-2.7.3/bin/python \
+	${htseq_count} \
 	-m union \
 	-s no \
 	- \
 	${annotationGtf} | \
-head -n -5 \
-> ${txtExpression}
+	head -n -5 \
+	> ${txtExpression}___tmp___;
+then
+	echo "Gene count succesfull"
+	mv ${txtExpression}___tmp___ ${txtExpression}
+else
+	echo "Genecount failed"
+fi
+
+rm ${sortedBam%bam}byName
 
 echo "Finished!"
 </#noparse>

compute4/RNA-seq_quantify_gene_level/protocols/MakeExpressionTable.ftl

@@ -1,20 +1,25 @@
-#MOLGENIS walltime=6:00:00 nodes=1 cores=1 mem=4
+#MOLGENIS walltime=24:00:00 nodes=1 cores=1 mem=6
 
 #FOREACH mergedStudy
 mkdir -p ${expressionFolder}
 
 rm -f ${expressionFolder}/fileList.txt
 
-<#assign samples=sample?size - 1>
-<#list 0..samples as i>
-  echo -e "${sample[i]}\t${txtExpression[i]}" >> ${expressionFolder}/fileList.txt
+<#assign runs=run?size - 1>
+<#list 0..runs as i>
+  echo -e "${run[i]}\t${txtExpression[i]}" >> ${expressionFolder}/fileList.txt
 </#list> 
 
-${JAVA_HOME}/bin/java \
+if ${JAVA_HOME}/bin/java \
 	-Xmx4g \
 	-jar ${processReadCountsJar} \
 	--mode makeExpressionTable \
 	--fileList ${expressionFolder}/fileList.txt \
 	--annot ${geneAnnotationTxt} \
-	--out ${expressionTable}
-
+	--out ${expressionTable}___tmp___
+then
+	echo "table create succesfull"
+	mv ${expressionTable}___tmp___ ${expressionTable}
+else
+	echo "table create failed"
+fi