Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

PCR for GAPDH (glyceraldehyde-3-phosphate dehydrogenase) detection

SC Sonia Carmen Lopo Costa
JF Jéssica de Souza Freitas
FC Fábio Santos Carvalho
MP Maria Julia Salim Pereira
MC Matheus Dias Cordeiro
AF Adivaldo Henrique da Fonseca
MJ Márcia Mariza Gomes Jusi
RM Rosangela Zacarias Machado
AM Alexandre Dias Munhoz
ask Ask a question
Favorite

To verify DNA integrity and the presence of potential inhibitors, negative samples were subjected to PCR for detection of the GAPDH gene using primers described by Birkenheuer et al. [38] (Table ​(Table1).1). In the PCR reactions, a final volume of 25 μl was used, composed of 5 μl of genomic DNA, 10 × reaction buffer, 2.0 mM of MgCl2, 0.2 mM of each dNTP, 0.4 μM of each primer, 1.25 U of Taq polymerase, and ultrapure water until the final volume was reached. The amplification protocol used consisted of an initial denaturation step at 95 °C for 5 min, followed by 40 cycles at 94 °C for 30 s for denaturation, annealing at 52 °C for 1 min, extension at 72 °C for 1 min, and final extension at 72 °C for 5 min [39].

Copyright and License information: The Author(s) ©2021
Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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.

post Post a Question
0 Q&A