The Desired Length of the Library Insert was Influenced by the Degree of mRNA Purification in Poplar
1 State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 51 Hexing Road, Harbin 150040, China
2 Children’s Hospital of Harbin, 57 Youyi road, Daoli District, Harbin 150010, China
3 Fuxing Hospital of Capital Medical University, 20 Fuxingmen srteet, Xicheng District, Beijing, 10069, China
Plant Gene and Trait, 2017, Vol. 8, No. 6 doi: 10.5376/pgt.2017.08.0006
Received: 27 Aug., 2017 Accepted: 15 Oct., 2017 Published: 29 Oct., 2017
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Preferred citation for this article:
Wang L.N., Ya F.L., and Wang L.F., 2017, The desired length of the library insert was influenced by the degree of mRNA purification in Popla, Plant Gene and Trait, 8(6): 61-66 (doi: 10.5376/pgt.2017.08.0006)
The typical workflow of an RNA-seq assay involves the extraction and further purification of mRNA. The size of the target DNA fragments in the final library is a key parameter for RNA-Seq library construction. In our experiment, we found that fragmentation is influenced by the purification of mRNA and leads to different insert sizes in the transcriptome libraries. This study compared many purification methods after extraction mRNA using magnetic silica beads. To assess the quality of the mRNA obtained from these options, the size of library was analyzed on the Agilent 2100 Bioanalyzer to measure whether the library is constructed successfully. Results of the best purification method could thoroughly remove the rRNA, tRNA and other impurities to obtain complete, high-purity mRNA molecules. The discovery of this phenomenon, the summary of the rules and the related purification reagents ratio all these can help us further exploited RNA-seq protocols.
mRNA; RNA-Seq; Transcriptome library; Fragmentation