GSM7794360
Centrifuge plates at 100xg for 3 mins, remove supernatant. Wash 1X with 100uL PBS, aspirate. Add 100uL lysis buffer, pipet up and down and freeze at -80C. Libraries were constructed using a modified SmartSeq2 protocol.RNA lysate were purified with Dynabeads mRNA Direct Purification kit (Life Technologies),followed by reverse transcription with Maxima Reverse Transcriptase (Life Technologies) and whole transcription amplification (WTA) with KAPA HotStart HiFi 2x ReadyMix (Kapa Biosystems).WTA products were purified with Ampure XP beads (Beckman Coulter). RNA-Seq libraries were constructed using Nextera XT DNA Library Preparation kit (Illumina). Libraries were sequenced with Nextseq500(75cycles) paired-end and the following parameters : paired End: Index1 8, Index2 8, Read1 38, Read2 38