GSM8758206: hippocampus_Mice_IPA1; Mus musculus; RNA-Seq
(1) Nanodrop detects RNA purity (OD260/280), concentration, and whether the nucleic acid absorption peak is normal; (2) Agilent 2100 accurately detects the integrity of RNA, and the detection indexes include: 28S/18S, the presence or absence of uplift of the graph baseline, and 5S peaks. (1) Enrichment of eukaryotic mRNA with magnetic beads with Oligo (dT); (2) Random interruption of mRNA by adding Fragmentation Buffer; (3) Synthesize the first cDNA strand using mRNA as a template with six-base random primers (random hexamers), then add Buffer, dNTPs, RNase H and DNA polymerase I to synthesize the second cDNA strand and purify the cDNA using AMPure XP beads; (4) The purified double-stranded cDNA was then subjected to end repair, addition of A-tail and ligation of sequencing junctions, followed by fragment size selection using AMPure XP beads; (5) Finally, the cDNA library was obtained by PCR enrichment.  Translated with DeepL.com (free version)