scripts/STARlong_MatchAnnot_commands_mapAs2ToGenome.txt

Version 1

#P. citri all data (VF+MF susphire and Edinburgh all Pcitri samples)

##mapping assembled transcripts to genome
# the genome is 486392826 nt long and has 17212 scaffolds, therefore genomeChrBinNbits was set to 15 [log2(486392826/17202)=14.8]

#mkdir star_genome_index
#
#STARlong --runThreadN 32 --runMode genomeGenerate \
#--genomeDir ./star_genome_index \
#--genomeFastaFiles ./input/Planococcus_citri_Pcitri.v1.scaffolds.fa \
#--sjdbGTFtagExonParentTranscript Parent \
#--sjdbGTFfile ./input/Planococcus_citri_Pcitri.v1.gff3 \
#--genomeChrBinNbits 15 \
#--limitGenomeGenerateRAM=220000000000

cp -avr /DATA/markop/Pcitri_rnaspades/star_genome_index .

mkdir STAR_mapSpadesTRtoGenome

#mapping to soft filtered transcriptome (soft_filtered_transcripts.fasta)
#parameters same as for the potato tr mapping but export BAM instead of SAM file
STARlong \
--runMode alignReads \
--outSAMattributes NH HI NM MD \
--outFilterType BySJout \
--outFilterMultimapNmax 20 \
--alignSJoverhangMin 8 \
--alignSJDBoverhangMin 1 \
--outFilterMismatchNmax 999 \
--outFilterMismatchNoverReadLmax 0.08\
--alignIntronMin 20 \
--alignIntronMax 1000000 \
--alignMatesGapMax 1000000 \
--outSAMtype BAM SortedByCoordinate \
--limitBAMsortRAM 100000000000 \
--runThreadN 28 \
--genomeDir ./star_genome_index \
--readFilesIn ./output/soft_filtered_transcripts.fasta \
--outFileNamePrefix ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong. \
--seedPerReadNmax 100000 \
--seedPerWindowNmax 1000 \
--seedSearchLmax 30 \
--seedSearchStartLmax 30 \
--alignTranscriptsPerReadNmax 100000 \
--alignTranscriptsPerWindowNmax 10000 \
--scoreGapNoncan -20 \
--scoreDelOpen -1 \
--scoreDelBase -1 \
--scoreInsOpen -1 \
--scoreInsBase -1 \
--chimMultimapNmax 20 \
--chimSegmentMin 100 

#index bam file
/DATA/majak/samtools-1.6/samtools index ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sortedByCoord.out.bam


#convert bam to sam for MatchAnnot
/DATA/majak/samtools-1.6/samtools view -@ 28 -O SAM -o ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sortedByCoord.out.sam ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sortedByCoord.out.bam
sort -k 3,3 -k 4,4n ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sortedByCoord.out.sam > ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sorted2.out.sam
rm ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sortedByCoord.out.sam

#gff3 to gtf
gffread ./input/Planococcus_citri_Pcitri.v1_mod.gff3 -T -E -F -O --gene2exon -o ./input/Planococcus_citri_Pcitri.v1.gtf

#MatchAnnot
python /home/administrator/Software/MatchAnnot/matchAnnot.py \
--gtf=./input/Planococcus_citri_Pcitri.v1.gtf \
--format=alt \
./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sorted2.out.sam > ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sorted2.out.matchAnnot.txt

#parsing the matchannot results: EvigenetrID  DMgeneID DMtrID exon_match match_score
grep "result:" ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sorted2.out.matchAnnot.txt | tr -s ' ' | awk -F'[ ]' '{print $2, $3, $4, $6, $8}' > ./STAR_mapSpadesTRtoGenome/PcSpadesTRsofttoGenome_STARlong.Aligned.sorted2.out.matchAnnot.parsed.txt