Decreasing miRNA sequencing bias using a single adapter and circularization approach

被引:42
作者
Barberan-Soler, Sergio [1 ]
Vo, Jenny M. [1 ]
Hogans, Ryan E. [1 ]
Dallas, Anne [1 ]
Johnston, Brian H. [1 ]
Kazakov, Sergei A. [1 ]
机构
[1] SomaGenics Inc, Santa Cruz, CA 95060 USA
关键词
miRNA-seq; Sequencing bias; Small RNA sequencing; Small RNA library preparation; RNA LIBRARY PREPARATION; MICRORNAS; LIGASE;
D O I
10.1186/s13059-018-1488-z
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The ability to accurately quantify all the microRNAs (miRNAs) in a sample is important for understanding miRNA biology and for development of new biomarkers and therapeutic targets. We develop a new method for preparing miRNA sequencing libraries, RealSeq (R)-AC, that involves ligating the miRNAs with a single adapter and circularizing the ligation products. When compared to other methods, RealSeq (R)-AC provides greatly reduced miRNA sequencing bias and allows the identification of the largest variety of miRNAs in biological samples. This reduced bias also allows robust quantification of miRNAs present in samples across a wide range of RNA input levels.
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页数:9
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