So I'm basically getting these 'successfully assigned reads" of around 30 - 45%. When I did STAR I got alignment reads of like 70-80% which is really good. I know that FeatureCounts counts the amount of reads (achieved in STAR) appeared to overlap with known genes. And so it's normal that it's lower than the percentage I got for the STAR alignment. However, I don't really get how it gets this low. Is there maybe a fault that I could've prevented while doing fastqc? Like some trimming or something? Would it then increase in percentage? (Assigned reads). And secondly, if I don't need to do anything. What can I give as an explanation for this low percentage? Can I just proceed with my analysis?
Load annotation file /mnt/storage/data/resources/genomes/hg38/hg38.ref ... ||
|| Features : 831683 ||
|| Meta-features : 28278 ||
|| Chromosomes/contigs : 367 ||
|| ||
|| Process BAM file C1.bam... ||
|| Single-end reads are included. ||
|| Assign reads to features... ||
|| Total reads : 35409751 ||
|| Successfully assigned reads : **13794555 (39.0%)** ||
|| Running time : 0.47 minutes ||
|| ||
|| Process BAM file C2.bam... ||
|| Single-end reads are included. ||
|| Assign reads to features... ||
|| Total reads : 33118524 ||
|| Successfully assigned reads : **14166283 (42.8%)** ||
|| Running time : 0.45 minutes ||
|| ||
|| Process BAM file S1.bam... ||
|| Single-end reads are included. ||
|| Assign reads to features... ||
|| Total reads : 31476163 ||
|| Successfully assigned reads : **13113126 (41.7%)** ||
|| Running time : 0.42 minutes ||
|| ||
|| Process BAM file S2.bam... ||
|| Single-end reads are included. ||
|| Assign reads to features... ||
|| Total reads : 38025040 ||
|| Successfully assigned reads : **12831852 (33.7%)** ||
|| Running time : 0.51 minutes ||
|| ||
|| Read assignment finished. ||
|| ||
|| Summary of counting results can be found in file "all.counts.summary"
Cross-posted
https://www.biostars.org/p/9547248/