RNA-Seq of mus musculus:tongue
Total RNA was isolated from tongue tissue and purified. The mRNA containing PolyA(polyadenylate) was specifically captured by using oligo(dT) magnetic beads (Dynabeads Oligo(dT), cat.25-61005, Thermo Fisher, USA). The captured mRNA was fragmented by NEBNextR Magnesium RNA Fragmentation Module (NEBNextR Magnesium RNA Fragmentation Module, cat. E6150S, USA) and synthesized into cDNA by reverse transcriptase. Then E. coli DNA polymerase I (NEB, cat.m0209, USA) and RNase H (NEB, cat.m0297, USA) were used for double-stranded synthesis. The composite double-stranded DNA and RNA were then converted into DNA double-stranded. The double strands were digested with UDG enzyme (NEB, cat. M0280, MA, US), pre-denatured, denatured, annealed and extended by PCR. The average insert size for the final cDNA library was 30050 bp. At last, we performed the 2150bp paired-end sequencing (PE150) on an illumina Novaseq 6000 (LC-Bio Technology CO., Ltd., Hangzhou, China) following the vendor's recommended protocol.