3T3_met40
The 40% methanol-fixed cells were mixed with 1l 10mM dNTP and 1l 100M oligo-dT30VN and incubated at 55C for 5min, then placed on ice immediately for 2min to facilitate RNA denaturation. Next, reverse transcription was performed by adding 6l RT mix. Full-length transcriptome amplification and quality control. After reverse transcription, cells were further lysed by adding 5l Proteinase K and incubating at 56C for 60 min and 95C 10 min. Then, the PCR reaction was performed after added 25l PCR buffer. PCR product was purified by DNA Clean & ConcentratorTM-5 kit.