GSM6600059: MEF_creAF10_rep2; Mus musculus; RNA-Seq
RNA was harvested in TRIzol and extracted with chloroform. The aqueous phase was isolated after spinning the samples at 21xg for 15 min at 4°C. 0.53 volumes of ethanol was added, mixed, and spun through Qiagen RNeasy column at 21xg for 30 seconds. DNA was digested on the column with 10 μL RNAse free DNase mixed with 70 μL RDD buffer (Qiagen, 79254) for 30 min at room temperature. The RNA was washed with 500 μL of buffer RW1 and RPE, each. RNA was eluted with 30 μL of water. 2 ug of total RNA was used for the construction of sequencing libraries. RNA libraries were prepared using TruSeq RNA Sample Prep Kit V2 (Illumina, RS-122-2002) following the manufacturer's instructions.