Reverse transcription PCR was carried out as follows: The amplification profile was 95℃ for 5 min(denaturation), followed by 30 cycles at 95℃/30 s; 56℃/30 s[GNAQ] or 55℃/30 s[COL1, OSX] or 60℃/30 s[RUNX2, OPN]; 72℃/1 min(Annealing). Lastly, a final extension step was performed for 5 min at 72℃. Quantitative PCR was carried out as follows: after an initial 5-minute enzyme activation step at 95℃, 40 amplification cycles were run, each consisting of 95℃ for 15 s and 60℃ for 30 s.
Copyright: Content may be subjected to copyright.
How to cite:
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
Seo, S(2022). Reverse transcription PCR and quantitative PCR analysis. Bio-protocol Preprint. bio-protocol.org/prep1796.
Choi, J., Lee, Y. S., Shim, D. M., Lee, Y. K. and Seo, S. W.(2021). GNAQ knockdown promotes bone metastasis through epithelial-mesenchymal transition in lung cancer cells. Bone & Joint Research 10(5). DOI: 10.1302/2046-3758.105.BJR-2020-0262.R3
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this
article to respond.
0/150
Tips for asking effective questions
+ Description
Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.
Spinning
Post a Question
0 Q&A
Spinning
This protocol preprint was submitted via the "Request
a Protocol" track.