GSM3502766
GSM3502766: wt_45_I; Candida albicans; RNA-Seq
RNA was isolated using Trizol and glass bead lysis after which RNA was precipitated and DNase treated followed by a column purification step. 5μg DNase-treated total RNA was subjected to mRNA purification using Dynabeads mRNA purification kit (Invitrogen). Remaining rRNA contamination was checked in the bioanalyzer using Agilent RNA 6000 Pico kit (Agilent). Purified mRNA was subjected to fragmentation using the NEBNext® Magnesium RNA Fragmentation Module (New England Biolabs) and purified with RNeasy Plus Mini kit (Qiagen). Then, first-strand cDNA was synthesized with 50 ng/μl random hexamer primers using SuperScript ® III First-Strand Synthesis System for RT-PCR (Invitrogen). dNTPs were eliminated by purifying with MiniQuickSpin DNA Columns and subjected to second strand cDNA synthesis followed by clean-up with Minielute Reaction cleanup kit (Qiagen). cDNA concentration was measured using Quant-iT Picogreen dsDNA Reagents (Invitrogen) in a NanoDrop Fluorospectrophotometer ND-3300 (Thermo Fisher). Library preparation was done using the KAPA DNA Library Preparation Kit (Kapa Biosystems) according to the manufacturer's instructions.
GEO
GSM3502766
GEO Accession
GSM3502766