TurboCapture 96 mRNA Kit (5)

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Publication protocol

RT-QPCR analysis: The mRNAs were isolated from SGC-7901 cells using TurboCapture 96 mRNA Kit (QIAGEN, USA) 48 h post-transfection, and then were submitted to a 25 μl PCR reaction in the presence of 12.5 μl of 2× Master Mix, 1 μl of each forward and reverse primers mix (10 μM each), 0.5 μl of 50× SYBR Green I and 4 μl mRNA as template. The PCR mixtures were first subjected to reverse transcription for 30 min at 42 °C and initially denatured for 5 min at 95 °C, and then to 45 cycles of amplification with the following cycling condition: 20 s at 95 °C, 30 s at 58 °C, and 30 s at 72 °C. The primer pairs for each gene were designed with Primer Premier 5.0 software; glyceraldehyde-3-phosphate dehydrogenase served as an internal control for PCR. Sequences of all the primers (Biomics Biotech, China) are shown in Table 2.

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