GSM4928860: Chinese Red Steppes 4 [RC4]; Bos taurus; RNA-Seq
Total RNA was isolated from longissimus dorsi muscles using Trizol Reagent (Invitrogen, USA) according to the manufacturer's instructions. Total RNA was treated with DNase I (NEB, Beijing, China), extracted with phenol-chloroform, and precipitated with ethanol. The quality and quantity of total RNA were determined using an Agilent 2100 Bioanalyzer (Agilent technologies, Palo Alto, CA). The mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. 1.5 µg of mRNA from each sample was used to construct six cDNA libraries for sequencing. The mRNA was treated with a Thermomixer (Eppendorf AG, Hamburg, Germany) to generate fragments with an average size of 200 bp (200 ± 25 bp) for the paired-end libraries. The fragmented mRNA was then used as templates for synthesizing the first-strand cDNA. The double-stranded cDNAs were purified and ligated to adaptors for Illumina paired-end sequencing. Library concentration was quantified by qPCR and a Qubit® 2.0 Flurometer (Life Technologies, CA, USA), and the insert size was checked on an Agilent Bioanalyzer 2100 system (Agilent Technologies, Palo Alto, CA).
GEO Accession
GSM4928860