title: GSM8545405: MDA-MD-231_Leu2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated using Trizol reagent (Invitrogen), according to manufacturer's instructions. Rapid gel extraction was used, followed by overnight precipitation at −20 °C for RNA extraction from the polyacrylamide gels. Ribosomal RNA depletion was not performed. Prepared RPF libraries were analyzed on a 2100 Bioanalyzer using a 7500 chip (Agilent, Santa Clara, CA) and quantified by qPCR (Cat.no. 07960140001, Roche, Switzerland) and subsequently diluted to a 10 nM concentration. Next, the libraries were pooled and loaded with 1.5 pM on a NextSeq550Dx system (RUO mode) and sequenced with 75-base single-read using a 75 Cycles High Output Kit v2.5 (Illumina Inc., San Diego). Total RNA was isolated using Trizol reagent (Invitrogen), according to manufacturer's instructions. Rapid gel extraction was used, followed by overnight precipitation at −20 °C for RNA extraction from the polyacrylamide gels. Ribosomal RNA depletion was not performed. Prepared RPF libraries were analyzed on a 2100 Bioanalyzer using a 7500 chip (Agilent, Santa Clara, CA) and quantified by qPCR (Cat.no. 07960140001, Roche, Switzerland) and subsequently diluted to a 10 nM concentration. Next, the libraries were pooled and loaded with 1.5 pM on a NextSeq550Dx system (RUO mode) and sequenced with 75-base single-read using a 75 Cycles High Output Kit v2.5 (Illumina Inc., San Diego).