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Quantitative Real-Time PCR

YW Yao Wang
JL Jing-jing Li
HB Hong-jun Ba
KW Ke-feng Wang
XW Xi-zhi Wen
DL Dan-dan Li
XZ Xiao-feng Zhu
XZ Xiao-shi Zhang
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Total cellular RNA was isolated with TRIzol reagent (Invitrogen). Samples were treated with DNase using the RNase-free DNase Set (Qiagen) during the total RNA isolation. First-strand complementary DNA (cDNA) was synthesized using the cDNA Synthesis kit (Thermo Fisher Scientific) according to the manufacturer's instructions. ABI prism 7900-HT sequence detection system (96-well, Applied Biosystems) was used to perform quantitative real-time PCR (RT-PCR) analysis. For RT-PCR, the following primers were used:

c-FLIPL: 5'-AGAACCTGGCTGCACCTAAC-3'(forward),

5'-GAGAAGGTCAAACCGCCTCA-3'(reverse).

GAPDH: 5'-CGAGATCCCTCCAAAATCAAGTGGGG-3'(forward),

5'-ACACGTTGGCAGTGGGGACAC-3'(reverse).

Each experiment was repeated three times and the relative expression of c-FLIPL was normalized to GAPDH expression.

Copyright and License information:  ©2019 Wang, Li, Ba, Wang, Wen, Li, Zhu and Zhang.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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