jec 2dRNA-seq
The mRNA Capture Beads (Yeasen,Item number: 12301E596) were added to 1ug total RNA to bind mRNA to magnetic beads, beads are washed and suspended in Beads Wash Buffer (Yeasen,item No.12301ES96) and Tris Buffer (Yeasen,item No.12301ES96).Synthesis of double-stranded cDNA.The first strand cDNA was synthesized by reverse transcription of the fragmented mRNA, followed by the second strand cDNA.The Adapter (Yeasen,item No.12414ES02/12415ES02) was added to the synthesized cDNA product.After the ligation,25 l(0.25x)VAHTS DNA Clean Beads.The purified connectors were amplified by PCR.After amplification,45)VAHTS DNA Clean Beads were used to purify the PCR products.Libraries were then examined using an Agilent 2100 Bioanalyzer and a Qubit 3.0Fluorometer (Invitrogen).