GSM8425668: Neuro-2a cells, shArf1-1-3; Mus musculus; RNA-Seq
RNA was hanvested using Trizol. Total RNA was used for the construction of sequencing libraries. In this project, mRNA with polyA structure in total RNA was enriched by Oligo(dT) magnetic beads, and ion interruption was used to break the RNA into fragments of about 300bp in length. The length of 300bp was chosen because the length of the junction is fixed, if the length of the interrupted fragment is shorter, it will lead to a high proportion of the junction sequence, which will reduce the proportion of valid data; if the length of the interrupted fragment is longer, it is not conducive to the generation of clusters in the process of on-line sequencing. The first strand of cDNA was synthesised using RNA as a template with 6-base random primers and reverse transcriptase, and the first strand cDNA was used as a template for the second strand cDNA synthesis.