Tools for qRT-PCR data analysis

Question

[mohammedtoufiq91]

Hi,

I have the qRT-PCR data and I would like to analyse this data from the Normalisation till the downstream analysis. Basically I would like to perform the calculation of DCt, DDCt, and 2^(-DDCt) including some of the plots. Could you kindly assist me with this.

Thank you,
Toufiq

Answer 1

[ivivek_ngs]

Here you go, but try to look through a simple google search. You will find your answer. Providing below, ways, tutorials and available tools and software list available to perform what you need to do your own analysis. Good luck!

Analysis1

Tutorial 1

Tools survey article

Software lists

Answer 2

[l.souza]

This is the usual RT-qPCR analysis workflow I use:

1. Retrieve the Rn values and input at Miner to get CTs, reaction efficiency and some statistics;

2. Load the Miner info into REST 2009 withou choosing the reference genes;

3. Load the preliminar results into geNorm to pick up the best reference genes;

4. Back to REST 2009 and check the reference genes selected from geNorm;

In my lab, we have been using this protocol with a great success for some years.

Regards,

Lucas

[mohammedtoufiq91]

Thank you for the inputs.