Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Real-time PCR

HL Hui Li
DM Defang Meng
JJ Jieting Jia
HW Hua Wei
request Request a Protocol
ask Ask a question
Favorite

Total RNA was reversely transcribed to cDNAs using the HiScript II Q RT SuperMix (Vazyme Biotech, Nanjing, China). Later, PCR was implemented on the SYBR Green Master Mix and the StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA). Relative level was calculated by the 2–ΔΔCt method, with GAPDH as the internal reference. Gene-specific primers (Genscript Biotech, Nanjing, China) are listed as below. 5′-GCACTTCACCGCGACTGTTA-3′ as the PGLYRP2 forward primer sequence and 5′-TTATTGGAGGTCTGTGGCTGG-3′ as the reverse primer sequence resulted in a PCR product of 103 bp. 5′-TCAGTGGTGGACCTGACCTG-3′ as the GAPDH forward primer sequence and 5′-TGCTGTAGCCAAATTCGTTG-3′ as the reverse primer sequence resulted in a PCR product of 244 bp.

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.

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