GSM3143852: ILC3-PBS-rep-2; Mus musculus; RNA-Seq
Total RNA was extracted using Trizol reagent (Invitrogen).Total RNA sample is digested by DNaseІ (NEB), and purified by oligo-dT beads (Dynabeads   mRNA purification kit, Invitrogen) Poly (A)-containing mRNA were fragmented into 130bp   with First Strand buffer. First-strand cDNA is generated by N6 primer, First Strand Master Mix and   Super Script II reverse transcription(Invitrogen) (Reaction condition：25℃ for 10 min；42℃ for 40   min；70℃for 15 min). Then add Second Strand Master Mix to synthesize the second-strand cDNA   (16℃ for 1h). Purified the cDNA with Ampure XP Beads (AGENCOURT), then combine with End   Repair Mix, incubate at 20℃for 30min. Purified and add A-Tailing Mix, incubate at 37℃ for 30   min. Then combine the Adenylate 3'Ends DNA, Adapter and Ligation Mix, incubate the ligate   reaction at 20℃for 20 min. Several rounds of PCR amplification with PCR Primer Cocktail and   PCR Master Mix were performed to enrich the cDNA fragments. Then the PCR products are   purified with Ampure XP Beads (AGENCOURT).
GEO Accession
GSM3143852