GSM6318190: E14, WT, primed, replicate 2, PRO-seq; Mus musculus; OTHER
Nuclei were isolated using Dounce homogenizer (1 million cells per mL) and nuclear run-on was performed with all four biotin-NTPs. RNAs were extracted by Trizol LS (Ambion) and fragmentated by base hydrolysis. From the fragmentated RNAs, biotin RNAs were enriched by streptavidin beads. The biotin RNAs were enriched twice more in each 3' and 5' RNA adaptor ligation processes. Reverse transcription, PCR amplification, and library size selection were performed to obtain 140 ~ 350bp, 5ng/uL libraries.