nascent RNA from pSer2-RNAPII-IP followed by Mnase digestion (mNET-seq) nascent RNA was used for library preparation with NEBNext Multiplex Small RNA Library Prep Set for Illumina Set 1, Set 2, Index Primers 1–48 and Multiplex Compatible Set1 (E7300). The libraries were amplified with 16 PCR cycles depending on the input RNA. The concentration and size distribution of the libraries were determined on a Fragment Analyzer using the HS Small Fragment (50-1,500 bp; Agilent Technologies) and NGS Fragment High Sensitivity Analysis Kit (1-6,000 bp; Agilent Technologies). The libraries were sequenced on the NextSeq2000 Illumina platform for 75 cycles. Base calling was performed using Illumina's BaseSpace platform mNET-seq