Hi,
I'm writing as I have obtained an RNAseq dataset and have commenced mapping with STAR. I have chosen to add the option of clipping the reads using the added option: '--clip3pNbases 75', to clip 75 bases from the end of the read. I am working with an RNAseq dataset where reads are 150bp. I have used the following code:
STAR --genomeDir ~/RNAseq/Resources/STAR_idx/ \
--runThreadN 6 \
--readFilesCommand zcat \
--readFilesIn ~/RNAseq/DRG_genewiz2/00_fastq/${x}_R1_001.fastq.gz ~/RNAseq/DRG_genewiz2/00_fastq/$$
--outFileNamePrefix ~/RNAseq/DRG_genewiz2/STAR_output_genewiz2/${x} \
--outTmpDir ~/RNAseq/temp \
--outSAMtype BAM SortedByCoordinate \
--quantMode GeneCounts \
--clip5pNbases 0 \
--clip3pNbases 75
done
However when I open the Log.final.out file for my mapping it lists average mapped length as ~149. To me this sounds like the clipping hasn't occurred? Perhaps I am misunderstanding something? I hope someone can clarify.
1 answer
I realized that the average mapped length of ~149 corresponds to the clipping having occurred. Had the reads not been clipped, the average mapped length would have been 300 when sequencing is carried out with 150bp reads.
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Have you looked at the resulting BAM files to see if the alignments only show 75 bp?