# 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.

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

`=D6-E6`

• Press Enter.

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

`=F6-G6`

• Press Enter.

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

`=2^(-H6)`

• Press Enter.

• 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.

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)`

• Press Enter.

• Double-click on the Fill Handle icon to fill the formula up to cell E11.

• Estimate the average of the three control values.
• Enter the following formula in the merged F6.

`=AVERAGE(E9:E11)`

• Press Enter.

• Select cell G6 and enter the following formula to get the value of ∆Ct. Input the Absolute Cell Reference for cell F6.

`=E6-\$F\$6`

• Press Enter.

• Double-click on the Fill Handle icon to fill the formula up to cell G11.

• Follow the same process complete all the calculations for the Gene of Interest (GOI) section.

• In cell O6, enter the following formula to get the value of the expression ratio. Ensure the Absolute Cell Reference for cells N6 and H6.

`=(\$N\$6^M6)/(\$H\$6^G6)`

• Press Enter.

• Double-click on the Fill Handle icon to fill the formula up to cell O11.
• You will get the values of the expression ratio for every sample.

Read More: How to Analyze Large Data Sets in Excel

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Soumik Dutta

Soumik Dutta, having earned a BSc in Naval Architecture & Engineering from Bangladesh University of Engineering and Technology, plays a key role as an Excel & VBA Content Developer at ExcelDemy. Driven by a profound passion for research and innovation, he actively immerses himself in Excel. In his role, Soumik not only skillfully addresses complex challenges but also demonstrates enthusiasm and expertise in gracefully navigating tough situations, underscoring his unwavering commitment to consistently deliver exceptional, high-quality content that... Read Full Bio

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