title: GSM8602309: NUN-Low-Exp2-RepB; Homo sapiens; OTHER
library_construction_protocol: For total RNA extraction :  cell pellets were resuspended in 1 mL of Trizol and RNA was extracted following manufacturer's instructions. For Nascent RNA extraction :  cell pellets were lysed in a hypotonic buffer, nuclei were purified on sucrose cushion, the chromatin fraction was then extracted using a high-salt high-urea concentration buffer (300mM NaCl, 1M Urea, 1% NP-40) and chromatin-bound nascent RNA was further extracted using Trizol according to manufacturer´s instructions. Total RNA or  Nascent RNA were processed using NEBNext Ultra II directional RNA library prep kit for Illumina (#E7760) using section 1 “Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module” or section 4 “Protocol for use with purified mRNA”, respectively.