GSM9885735: 66_1E4_AAV-miMSH3_rep2_7_days; Homo sapiens; RNA-Seq
Total RNA was isolated by lysing cells in 350 µL RLT Buffer (containing 1% 2-mercaptoethanol) and homogenized with the QiaShredder (Qiagen 79656), followed by extraction with the RNeasy Micro Kit (Qiagen 74004) according to manufacturer's instructions including the optional on-column DNase digestion (Qiagen 79254). RNA was eluted in RNAse-free water and RIN and concentration were measured using the Fragment analyzer 5300 (Agilent). Samples with RIN > 7 were further analyzed. Libraries were prepared with 300 ng of RNA input by the Biomek i7 Hybrid automation (Beckman Coulter) with the Universal Plus mRNA-Seq with NuQuant kit with Custom AnyDeplete (NuGen 0522-A01). Library preparation with polyA transcript selection followed the supplier protocol and libraries were amplified with 13 cycles.