RNA isolation, quantitative polymerase chain reaction, and bulk RNA-seq
This protocol is extracted from research article:
A transcriptional roadmap for 2C-like–to–pluripotent state transition
Sci Adv, May 29, 2020; DOI: 10.1126/sciadv.aay5181

Cellular RNA was isolated using Qiagen Allprep RNA/DNA Mini Kit (Qiagen, catalog no. 80204). Complementary DNA was prepared by using the SuperScript III First-Strand Synthesis System (Thermo Fisher Scientific, catalog no. 18080051) and quantitative reverse transcription polymerase chain reaction was performed by using the Fast SYBR Green Master Mix (Thermo Fisher Scientific, catalog no. 4385612). Relative quantification was performed using the comparative cycle threshold (CT) method normalized to Gapdh. Bulk RNA-seq sample was generated by the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB, catalog no. E7420S).

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