i am working on the expression of some growth factors, i want to know the accurate and best method to calculate the relative mRNA expression of a target gene (VEGF) among three groups (1 control, 2 vehicle, 3 treatment) by using CT values obtained by RT-PCR .
Dear Vinay, Good Morning.
In my experience there is a program on MS Excel called GENEX which will make your life easy for the calculations. You just need to copy and import the Well number, Ct value and the identifier and it will give you the analysis with a single mouse click. If you give me your mail address i can forward it to you. my email address is [email protected].
Best Regards.
You welcome, i have sent you GENEX and detailed email.
Good luck with your analysis.
Dear Zulfiqar Naqvi,
I am also working with Real Time PCR, and eager to know about this Application. Could you please send me too.
My mail Id is [email protected]
Hi all. Comaprative method, aka 2^-deltaCt method, is the most accurate for determining the mRNA expression in samples and for comparison among experimental groups. Look at this article
PMID: 11846609
Also, there are several web-based or downloadable free software that will enable to determine the most stable/accurate/suitable housekeeping gene. I found DataAssit from LifeTechnologies quite straightforward
http://www.lifetechnologies.com/us/en/home/global/forms/dataassist-software-registration.html
I recently published a step by step protocol for this in Methods in Molecular Biology - https://www.researchgate.net/publication/258638535_The_quantitative_real-time_polymerase_chain_reaction_for_the_analysis_of_plant_gene_expression?ev=prf_pub Hopefully this will help.
Article The Quantitative Real-Time Polymerase Chain Reaction for the...
Dear Zulfiqar
I also work in qPcr and I will be eager to try this application. I use qPcr relative to an expression curve.
best regards
my email address is [email protected]
Try the search function on RG and you will find a number of similar questions and many valuable answers.
https://www.researchgate.net/search.SearchPosts.html?query=qpcr&accountHitCount=128&topicHitCount=0&postHitCount=2139&pubHitCount=10828&jobHitCount=892&institutionHitCount=0&departmentHitCount=1&dbw=true
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