GSM3525826: WT-4weeks-rep3; Mus musculus; RNA-Seq
Freshly isolated livers were immediately frozen and stored in liquid nitrogen until use. mRNA was extracted using Trizol regent (Invitrogen) following the manufacturer's instructions. For mRNA-seq analysis, mRNA was isolated with Dynabeads® mRNA Purification Kits (Invitrogen), fragmented in 5x first strand buffer(Invitrogen) at 95°C for 5 min, and reverse-transcribed using SuperScript® II Reverse Transcriptase (Invitrogen). After the first strand was synthesized, Second Strand Buffer (Invitrogen), RNase H (New England Biolabs), dNTPs, and DNA polymerase I (TaKaRa) were added to the second-strand synthesis reaction for 2.5 h at 16°C, and cDNA was purified with Agencourt AMPure XP beads (Beckman). Libraries were constructed with a NEBNext DNA Library Prep Master Mix Set for Illumina (New England Biolabs), PCR products were purified using AMPure XP beads. RNA libraries were quantified by Qubit 1.0 (Invitrogen) and analyzed with an Agilent 2100 bioanalyzer(Agilent) for size distribution, and were sequenced with the Illumina Hiseq-2500 platform.
GEO Accession
GSM3525826