GSM9465379: VEH3; Mus musculus; RNA-Seq
Total RNA was extracted from the heart using TRIzol® Reagentaccording the manufacturer's instructions (Magen). RNA samples weredetected based on the A260/A280 absorbance ratio with a Nanodrop ND-2000 system (Thermo Scientific, USA), and the RIN of RNA was determinedby an Agilent Bioanalyzer 4150 system (Agilent Technologies, CA, USA). Onlyqualified samples will be used for library construction. Paired-end libraries were prepared using a ABclonal mRNA-seq Lib PrepKit (ABclonal, China) following the manufacturer's instructions. The mRNAwas purified from 1 μg total RNA using oligo (dT) magnetic beads followed byfragmentation carried out using divalent cations at elevated temperatures inABclonal First Strand Synthesis Reaction Buffer. Subsequently, first-strandcDNAs were synthesized with random hexamer primers and ReverseTranscriptase (RNase H) using mRNA fragments as templates, followed bysecond-strand cDNA synthesis using DNA polymerase I, RNAseH, buffer, anddNTPs. The synthesized double stranded cDNA fragments were then adapter-ligated for preparation of the paired-end library. Adaptor-ligated cDNA wereused for PCR amplification. PCR products were purified (AMPure XP system)and library quality was assessed on an Agilent Bioanalyzer 4150 system.Finally, sequencing was performed with an Illumina Novaseq 6000 /MGISEQ-T7 instrument.