Entering edit mode
3.0 years ago
arctic
▴
40
Hi all, I am relatively new to the field. I would be very grateful to have a second opinion on below options from someone with STAR experience (particularly in the context of AT or plant species).
Star [create genomic indices]
STAR \
--runMode genomeGenerate \
--genomeDir ...\
--genomeFastaFiles ./Arabidopsis_thaliana.TAIR10.dna.toplevel.fa \
--sjdbGTFfile ./Arabidopsis_thaliana.TAIR10.50.gtf \
--sjdbOverhang 74 \
\## My ReadLength-1
--genomeSAindexNbases 12
\## log2(GenomeLength)/2 - 1=12.41
Star [alignment]
STAR \
--quantMode GeneCounts \
--genomeDir ./ATGenoIndices \
--readFilesIn .... \
--outFileNamePrefix ... \
--outFilterMultimapNmax 20 \
--alignSJoverhangMin 8 \
--alignSJDBoverhangMin 8 \
--outFilterMismatchNmax 8 \
--alignIntronMin 35 \
--alignIntronMax 2000 \
\## 99.3% of introns in AT are below this size based on doi:10.3390/genes8080200
--alignMatesGapMax 100000 \
--readFilesCommand zcat
I have been trying to find the optimum settings using available literature, however the variety has been overwhelming. I eventually ended up pasting together above options from few references that appeared to be more in line with the analysis I am hoping to perform ( Which is simple DE analysis at gene level.).
Many thanks for your time and help in advance,