alias: E-MTAB-7090:H4T80A.Rep1_s
title: NextSeq 500 sequencing; Global gene expression analysis of H4T80A mutants vs WT H4 in Saccharomyces cerevisiae
design: Global gene expression analysis of H4T80A mutants vs WT H4 in Saccharomyces cerevisiae
library_construction_protocol: For each strain, three independent colonies were used to prepare samples of total RNA.  Wild-type H4 and otherwise isogenic T80A H4 cells were grown in yeast extract, peptone, glucose, adenine sulfate (YPDA) to an optical density at 660 nm (OD660) of 0.6.  Total RNA was isolated through hot-phenol extraction. RNA was further subjected to DNase I treatment (catalog number E1009-A; Zymo Research) and poly-A mRNA was purified using poly-T oligo-attached magnetic beads (NEXTflex Poly(A) Beads, catalog number NOVA-512981) according to the manufacturer's instructions. The quantity and purity of RNA were measured by using an Agilent high-sensitivity RNA screen tape system (catalog number 5067-5579; Agilent Technologies) and Qubit (Molecular Probes, Invitrogen).  Libraries were prepared for sequencing from 200 ng of polyA mRNA by using the NEXTflex RNA-seq kit (NOVA-512913, Illumina) according to the manufacturer's instructions. Samples were barcoded and combined at uniform molarity to create a single pool for sequencing.
identifiers.namespace: University of Portsmouth
identifiers.id: E-MTAB-7090:H4T80A.Rep1_s