GSM3478747: 4_901_6_13_17_Bx1; Homo sapiens; RNA-Seq
Snap frozen ectocervical biopsies were cut and placed into 350µl of RLT lysis buffer (QIAGEN) and 1% 2-mercaptogthanol (Sigma-Aldrich). The tissues were bead milled with a stainless-steel bead and Tissuelyser II (QIAGEN). Total RNA was extracted using RNeasy Micro Kit with DNase digestion (QIAGEN). RNA quantity and quality were assessed using Nanodrop 2000 Spectrophotometer and Agilent's 4200 Bioanalyzer Capillary electrophoresis. RNA quantity and quality were assessed using Nanodrop 2000 Spectrophotometer and Agilent's 4200 Bioanalyzer Capillary electrophoresis. 1ng of total RNA was used for mRNA amplifications using Clontech Smarter V4 chemistry (Takara Bio Inc). Amplified mRNA was fragmented and barcodes appended using Illumina's Nextera XT kits. Amplified libraries were validated using an Agilent 4200 Tapestation and quantified using a Qubit fluorimeter Libraries were normalized, pooled and clustered on an Illumina HiSeq 3000/4000 flowcell using an Illumina cBOT. Libraries were sequenced on an Illumina HiSeq 3000 system in 101-base single-read reactions with multiplexing to achieve approximately 20 million reads per sample.
GEO Accession
GSM3478747