mapping to transcriptome.fasta vs genome.fasta
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6 months ago
baibhu1234 ▴ 60

hello everyone, I am using nanopore DRS reads to find the m6A sites. I have mapped the reads to reference transcriptome (transcript.fasta from phytozome) so that we get the sites on the basis of transcripts. Now as i see the mapping bam file, all the reads (>99%) are mapped to + strand and very few to negative strand.

(for testing purpose i mapped to genome and found ~equal number of reads mapped to each strand). Also to check the strand of reference transcripts i check the gtf file, it shows equal number on both strands.

so my question is why are the reads mapped only to + strand when mapped to transcript.fasta? Is this have something to do with how the transcript.fasta is made from gtf? do they convert even the - strand transcritps to "+" or forward or sense or 5' to 3' strand?

mapping gtf transcriptome genome • 578 views
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Entering edit mode
6 months ago
Mensur Dlakic ★ 30k

All transcripts are shown in a + direction (a sense direction, if you will), regardless of their actual orientation in chromosomal DNA.

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