今天使用单端测序的 bam 文件进行表达值calling,命令如下所示:
rsem-calculate-expression
--alignments
-p 8
file.sorted.bam
/reference/rsem_ref/GRCh37/GRCh37
file
报错内容为: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!
检查以后才知道,虽然都叫bam,但是RSEM进行表达值calling的bam文件是比对到转录组的bam(transcriptome alignments),而非比对到基因组的bam(genomic alignments)。所以如果是比对到基因组的bam是不能用rsem直接计算表达值的,得转为fq再进行表达值计算。