今天使用單端測序的 bam 文件進行表達值calling,命令如下所示:
rsem-calculate-expression \
--alignments \
-p 8 \
file.sorted.bam \
/reference/rsem_ref/GRCh37/GRCh37 \
file
報錯內容為:The SAM/BAM file declares less reference sequences (25) than RSEM knows (196520)! RSEM can not recognize reference sequence name chr1! -tag XM" failed! Plase check if you provide correct parameters/options for the pipeline!
檢查以后才知道,雖然都叫bam,但是RSEM進行表達值calling的bam文件是比對到轉錄組的bam(transcriptome alignments),而非比對到基因組的bam(genomic alignments)。所以如果是比對到基因組的bam是不能用rsem直接計算表達值的,得轉為fq再進行表達值計算。