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The Degree of Purification of mRNA Influences the Fragmentation for Construction Transcriptome Libraries of Populus | Wang | Tree Genetics and Molecular Breeding

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The Degree of Purification of mRNA Influences the Fragmentation for Construction Transcriptome Libraries of Populus  

Li-Na Wang , Miao He , Yuanxue Zhao
State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 51 Hexing Road, Harbin 150040, China
Author    Correspondence author
Tree Genetics and Molecular Breeding, 2017, Vol. 7, No. 1   
Received: 25 May, 2017    Accepted: 27 Jun., 2017    Published: 12 Feb., 2018
© 2017 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract

The typical workflow of a RNA-seq assay involves the extraction and often further purification of mRNA from tissues; because rRNA reads are not informative it is best to reduce their levels. Fragmentation is essential factors and mostly library preparation protocols use for the detection of libraries, In our experiment the different reagents ratio were used to purify mRNA among those the highly purifies mRNA were used to construct transcriptome libraries. To assess the quality of the mRNA obtained from these methods, the cDNA libraries were analyzed on the Agilent 2100 Bioanalyzer. The option of 2.5 M LiCl binding buffer and 0.1 M LiCl elution buffer combined with 1% of LiDS could thoroughly remove the rRNA and other Impurities to obtain complete, high-purity mRNA molecules. The insights into molecular reactions that our framework allows can be further exploited to improve RNA-seq protocols, as we demonstrate experimentally.

Keywords
mRNA; RNA-Seq; Transcriptome library
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