Biases in Illumina transcriptome sequencing caused by random hexamer priming

被引:468
作者
Hansen, Kasper D. [1 ]
Brenner, Steven E. [2 ]
Dudoit, Sandrine [1 ,3 ]
机构
[1] Univ Calif Berkeley, Sch Publ Hlth, Div Biostat, Berkeley, CA 94720 USA
[2] Univ Calif Berkeley, Dept Plant & Microbial Biol, Berkeley, CA 94720 USA
[3] Univ Calif Berkeley, Dept Stat, Berkeley, CA 94720 USA
基金
美国国家卫生研究院;
关键词
RNA-SEQ; GENOME; BIOLOGY;
D O I
10.1093/nar/gkq224
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Generation of cDNA using random hexamer priming induces biases in the nucleotide composition at the beginning of transcriptome sequencing reads from the Illumina Genome Analyzer. The bias is independent of organism and laboratory and impacts the uniformity of the reads along the transcriptome. We provide a read count reweighting scheme, based on the nucleotide frequencies of the reads, that mitigates the impact of the bias.
引用
收藏
页数:7
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