GSM7434780: eWAT, AKO-1 1705_1; Mus musculus; RNA-Seq
Mettl14f/f and Mettl14Δfat male mice (8 weeks) were fed a HFD for 16 weeks and eWAT was dissected. Total RNA was extracted by RNeasy Mini Kit (Qiagen Cat#74104) and used for RNA-seq 1)mRNA enrichment: Oligo dT Selection or rRNA depletion;2)RNA fragment and reverse transcription:  Fragment the RNA and reverse transcription to double-strand cDNA (dscDNA) by N6 random  primer;3)End repair, add A tailing and adaptor ligation:The synthesized cDNA was subjected to end-repair  and then was 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA  fragments;4)PCR amplification:The ligation products were purified and many rounds of PCR amplification  were performed to enrich the purified cDNA template using PCR primer;5)Denature and  cyclization:Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and  DNA ligase;6) Sequencing on DNBseq platform.