How to Analyze Real time PCR Data? | Real Time PCR Gene Expression Fold Change Calculation

Welcome to my channel, "Learn Innovative with Shashi Bhushan Chauhan". In today's video, we delve into the nitty-gritty of Real-Time PCR Data Analysis and Fold Change Calculation, simplifying it for researchers and students alike.

In the realm of molecular biology, real-time PCR has become a cornerstone for quantifying gene expression. However, interpreting these results can often be challenging, particularly when calculating the fold-change gene expression data. This tutorial is designed to dispel those difficulties.

I will guide you through the process of calculating the fold change gene expression from real-time PCR data, using both Syber Green and Taqman Assays. This video addresses common obstacles faced by many research students, particularly when handling more than one control sample in their studies.

In this step-by-step tutorial, I'll show you how to calculate mean CT value, ΔCT value, ΔΔCT value, and finally, the fold change. I will demonstrate this process using Excel, making it a breeze to follow along.

In addition to this, I'll showcase how to represent your data visually, using graphs on Excel and the GraphPad Prism software, including creating an informative heat map.

This video is part of a series designed to make complex scientific processes accessible and understandable. So whether you're a seasoned researcher or a student starting in the field, this tutorial has something for you. If you find this video helpful, please like, share, and subscribe to support the channel.

Thank you for watching!

Key word for search:

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#Real Time QPCR Data Analysis Tutorial #How to Analyze Real-time PCR Data #How to Analyze Real-time PCR Data #How to Analyze Real-time PCR Data #Real Time PCR Analysis qPCR Terms

Time Code:
00:00 - Introduction
00:48 - Summary of all steps
01:03 - Calculation of Mean Ct value of each sample
02:20 - Calculation of delta Ct value
03:06 - Calculation of delta delta Ct value
04:20 - Fold Change Gene expression calculation
06:16 - Fold Change gene expression Graph in Excel
06:36 - Fold Change gene expression graph in Graph Pad Prism Software & Export

[grace-]

Very informative video, I was searching for this since long time...

[muneebmalik]

Excellent video, made my work so easier, thank you!

[shosugahara]

Thank you! This is absolutely what I wanted to know!

[rollymvogo]

Nicely, simply, and efficiently explained, very good video, Sir.

[josemariaespinosabernal]

Thank you very much for the video and especially for sharing the data

[snehasishnag]

Sir, so to get average and sd se values, i need to perform multiple qpcr? As for each qpcr, i am getting one del del cT value for control and one for the treated sample. Great video sir by the way

[Ghumakkad_kalaji]

Hi sir, Great video., Thank you very much

[sakkijarvenpolkka]

Thank you so much, very informative video! Saved me from hours of confusion and mislead!

[mgpqqle]

thank you so much, it's very clear

[amanidhiflaoui]

Thank you so much for these informative video!! It makes everything clear for me! Please keep share with us this kind of valuable information.

[grace-]

Thankyou soo much for such a informative video sir..

[grace-]

Somewhere I read that the treated group showed 20.42 higher fold change than control group and the corresponding log2 transformation was 4.37... sir how we get to know about this?

[leen_art]

Hello thank for the very informative video ❤ can you please explain how we can interpret the data afterwards like if the fold change increased how does that apply to the gene

[judith]

Nice video, how can i perform the calculation if my gene of interest is more expressed than my housekeeping gene? Thanks

[liadiafarida]

Thank you! You got new subscriber, sir!

[leen_art]

In these tests the treatment group and the control group are independent from each other correct?!

[ananyapalo]

Dear Sir, Can you please provide some paper references regarding this calculation.
So i can mentioned it in a paper

[chloev]

hi there, thank you for the great video. how would you go about doing this if you do not have control vs treated, instead you just want to compare genes of interest and housekeeping genes of different genotypes? Thanks!

[sheetal_soul]

nice explanation. I am trying real-time PCR with TaqMan probe with the manual master mix but I am not getting a curve graph for amplification curve. Can you explain real-time PCR with TaqMan probe (cy5)?

[selmasofiatorresvalles]

Thanks Shashi, excellent video. I have a question, is it necessary to have an endogenous control for each sample on the plate or can I have only one duplicate of endogenous control for all my duplicates of the same treatment? For example, one endogenous control duplicate for all my Treatment 1 samples, another endogenous control duplicate for all my samples from Treatment 2, and so on...