GSM4826435: mNSC-shNT; Mus musculus; RNA-Seq
Tree days after treatment, cells were harvested and total RNA were extracted with RNeasy® Micro Kit (Qiagen). The mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, a ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.