GSM3860470: HC2 [V_2]; Bos taurus; RNA-Seq
Total RNA extraction and purification of each sample was performed using the PicoPure RNA Isolation kit (Life technologies, Foster City, CA, USA) according to manufacturer´s instructions. Genomic DNA contamination was removed by performing an on-column DNA digestion using RNase-free DNase (Qiagen, Mississauga, ON, Canada). RNA was eluted in 11 µL of elution buffer and the RNA concentration was checked using a Qubit® 3 fluorometer (Life technologies), RNA integrity was evaluated using a Bioanalyzer 2100 (Agilent Technologies, CA, USA) with the use of RICH Pico Chips (Agilent Technologies). All samples analyzed had a RNA concentration ≥ 10 ng/µL an RNA integrity number (RIN) ≥ 7.7, and was considered adequate. The library preparation was performed using NEBNext Ultra II RNA Library Prep Kit for Illumina (New England, Biolabs, MA, EUA), according to manufacturer´s instructions