Total RNA (1 mg), isolated from cells using PerfectPure RNA Cultured Cell Kit (5 Prime) was reverse transcribed using random hexamers and Superscript II RNase H À Reverse Transcriptase (Invitro- gen). The reaction mixtures were diluted 20-fold, and 4.5 ml was then used as the template ...
For cDNA synthesis, 2 μg of total RNA was mixed with 1 mmol/L dNTP, 0.2 μg of a random primer, 20 U RNAsin, and 200 U M-MuLV reverse transcriptase in 20 μL of reverse transcription reaction buffer. The mixture was incubated at 42 °C for 60 min and subsequently heated at ...