Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Gene expression analysis by QPCR

MG Maarten H. Geurts
SG Shashank Gandhi
MB Matteo G. Boretto
NA Ninouk Akkerman
LD Lucca L. M. Derks
GS Gijs van Son
MC Martina Celotti
SH Sarina Harshuk-Shabso
FP Flavia Peci
HB Harry Begthel
DH Delilah Hendriks
PS Paul Schürmann
AA Amanda Andersson-Rolf
SL Susana M. Chuva de Sousa Lopes
JE Johan H. van Es
RB Ruben van Boxtel
HC Hans Clevers
request Request a Protocol
ask Ask a question
Favorite

Organoid RNA was reverse-transcribed and subjected to SYBR Greenbased quantitative real-time PCR (qPCR) using the forward and reverse primers described in Supplementary Table 5. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and hypoxanthine phosphoribosyltransferase (Hprt) were used as housekeeping genes. Relative gene expression levels were calculated as ΔCt values (Ct ‘target’ minus Ct ‘housekeeping gene’) and in most analyses compared between ‘sample’ and ‘reference’ to express fold change, i.e., 2-(ΔCt sample − ΔCt reference).

Copyright and License information: The Author(s) ©2023
Open Access This 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/.

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