How to Analyze qPCR Data in Excel (2 Easy Methods)

Method 1 – Analyze qPCR Data Through Double Delta Ct Method

Steps:

Input all the gene values accurately as shown in the image. We have to input all the experimental values of Gene Tested Experimental(TE) and Housekeeping Gene Experimental(HE) in columns B and C. Input all the control values of Gene Tested Control(TC) and Housekeeping Gene Control(HC) in columns D and E.

To calculate the value of ∆CTE, in cell F5, enter the following formula.

=B6-C6

Press Enter.

Analyse qPCR Data in Excel Through Double Delta Ct Method

To estimate the value of ∆CTC, enter the following formula in cell G5.

=D6-E6

Press Enter.

Analyse qPCR Data in Excel Through Double Delta Ct Method

Select cell H5 and enter the following formula to get the value of ∆∆Ct.

=F6-G6

Press Enter.

Analyse qPCR Data in Excel Through Double Delta Ct Method

To calculate the value of the Gene Expression Ratio or 2^(−∆∆Ct), enter the following formula in cell I5.

=2^(-H6)

Press Enter.

Analyse qPCR Data in Excel Through Double Delta Ct Method

Select the range of cell F6:I6.

Double-click on the Fill Handle icon to fill the formula up to row 10.
You will get the values of the Gene Expression Ratio for every sample.

Analyse qPCR Data in Excel Through Double Delta Ct Method

Interpretation of the Result

The value of the Gene Expression Ratio of 0.6507 in cell I5 means the gene samples in the tested conditions with respect to the control conditions all have been normalized to our housekeeping gene. You can consider it in the percentage term. 0.6507 means 65.07% gene expression in our tested condition with our control condition.

Method 2 – Analyze qPCR Using Pfaffl Method

Steps:

Input all the gene values accurately, as shown in the image. We have to input all the two sets of treated and control gene values of House Keeping Gene (HKG) and Gene of Interest (GOI). Both gene sets are denoted Ct-1 and Ct-2.

Complete all the calculations for the House Keeping Gene (HKG)
Take the mean value of both sets. For that, we are going to use the AVERAGE function.
Enter the following formula in cell E6.

=AVERAGE(C6:D6)