GSM3272711: Basal_Early_B; Mus musculus; RNA-Seq
Gastrocnemius muscles thawed for 1 minute at RT, then transferred to 2 ml TRI-reagent. Tissue was homogenized using disperser (IKA-T18) and then transferred to 1.7 ml tube and stand in RT for 5 minutes. 200 µL chloroform was added to each tissue sample for phase separation. Samples was then centrifuge for 15 minutes at 13,000 rpm at 4?c and transferred to tubes containing 500 µL Isopropanol. The upper phase was then isolated and stand at RT for 15 minutes and later centrifuged at 13,000 RPM at 4?c. Supernatant was removed and samples washed with 1ml of 75% cold EtOH and briefly vortexed. Sampled was then centrifuged for 10 minutes at 8,000 RPM at 4?c, and washed with 1ml of 75% cold EtOH and briefly vortexed. Pellet was dried, and later dissolved in 150 µL UPW. Samples was then incubated at 56 ?c for 10 minutes. RNA concentration was measured using Nanodrop. The TruSeq Stranded mRNA Sample Preparation kit (Illumina, San Diego, CA) was used. The libraries were prepared according to the manufacturer's protocol without further modifications