Hi,
I'm asking how can I use the gene counts of Candida albicans on R for the DE analysis? because every gene has 2 versions A and B because C. albicans is diploid. Can I sum the counts from A and B together? Ex:
for rca1:
C1_00020C_A 9309
C1_00020C_B 11
for control:
C1_00020C_A 6973
C1_00020C_B 128
Thanks, Zeinab
1 answer
How did you count reads mapping to genes? Some programs (e.g. featureCounts and HTSeq) by default do not count multimappers, so you would be under-estimating counts if you used these programs. My suggestion is to use Salmon or RSEM, then summarize the counts with tximport, using a "transcript to gene" map which maps both versions of each gene to one gene. See previous discussions:
C Albicans Genome Count Matrix Total Genes
Generate a counts matrix with paired-end non-stranded samples
P.S.: another possibility is to analyse allele-specific expression, but then I don't have a suggestion on how you should proceed.
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While that may be so I doubt that each one of your reads is able to tell them apart when you align them. In general people work with a single estimate of both copies for differential expression analysis.