GSM6589234: ESCA14T; Homo sapiens; RNA-Seq
Total RNAs (500 ng) from ESCC tissues and matched normal tissues were extracted by Trizol® Reagent (Thermo, USA) and treated with DNase I (NEB) to remove DNA before constructing the RNA-seq libraries. QIAseq FastSelect reagent was added to the RNA sample to fragment the RNA at 85°C for 6 minutes and then stepwise cool the reaction from 75°C to 25°C for 14 minutes. First- and second-strand cDNA was synthesized with random hexamer primers, treated with DNA End Repair Kit (Qiagen) to repair the ends, then modified with Klenow to add an A at the 3' end of the DNA fragments, and finally ligated to adapters. Purified dsDNA was further subjected to 11 cycles of PCR amplification. The libraries were quality controlled with Qubit (Thermo Fisher Scientific, USA) and Qsep100 (BiOptic, China)