Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Real-time qPCR

PP Paschalia Pantazi
MK Myrsini Kaforou
ZT Zhonghua Tang
VA Vikki M. Abrahams
AM Andrew McArdle
SG Seth Guller
BH Beth Holder
request Request a Protocol
ask Ask a question
Favorite

Real-time qPCR was performed using the Sso Advanced™ Universal Probes Supermix (Bio-Rad, 1725281) and the Taqman Gene Expression Assays GAPDH (Hs99999905_m1, VIC-MGB), XIST (Hs01079824_m1, FAM-MGB), and DDX3Y (Hs00190539_m1, FAM-MGB). The reactions were duplexed (GAPDH and gene of interest) and were run on an STEP-ONE real-time PCR system (Applied biosystems, 4376357). We mixed equal quantities of female and male placenta cDNA, which we run along with our samples, to serve as a calibrator, as it expressed both XIST and DDX3Y. The data were analyzed using the ΔΔCt method and the relative expression was calculated as 2−ΔΔCt.

Copyright and License information:  ©2022 The Authors
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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

tip 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.

post Post a Question
0 Q&A