基于RNA-seq技术的甘蓝型油菜新转录本的鉴定

摘要

In order to identify novel unknown transcripts in Brassica napus genome,total RNAs were extracted from leaves of Brassica napus,cDNA libraries were constructed,and sequenced using Illumina dual terminal sequencing Hiseq 2500 platform.Novel unknown transcripts were identified by bioinformatics analysis of high quality sequenced reads,and the results showed that 7 720 candidate novel transcripts were identified in the spacer region of known genes in Brassica napus genome.Fifteen newly identified transcripts were confirmed by RT-PCR and clone sequencing.%为了鉴定甘蓝型油菜基因组中新的未知转录本,以甘蓝型油菜叶片为材料提取总RNA,构建cDNA文库并使用Illumina双端测序Hiseq 2500平台进行测序.用生物信息学的方法分析高质量的测序reads来鉴定新的未知转录本.结果表明,在甘蓝型油菜基因组已知基因的间隔区共鉴定到7 720个候选新转录本,通过RT-PCR和克隆测序验证了15个新鉴定的转录本.