The Desired Length of the Library Insert was Influenced by the Degree of mRNA Purification in Poplar

Abstract

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.