How do I figure out the concentration of my DNA stock from the results I get from the QuantStudio 3D Digital PCR System?
If you include all of the necessary dilution factors into the AnalysisSuite software, the software will give you the copies/µL in your stock. There are 2 dilutions you need to take into account. The first is the dilution of the sample in the reaction. The second is the dilution of the stock that you make before adding it to the digital PCR reaction.
For example, say you want to add 1 µL of a sample that has been diluted 1:10 from the stock. If you add 1 µL of your sample to a 16 µL (final volume) reaction, the dilution factor of the sample is 1:16 or 1/16 = 0.0625. Since the stock has also been diluted 1:10 (0.1), you also need to factor this in. The final dilution factor to enter into the software is 0.0625 x 0.1 = 0.00625 (1:160). As you can see below, you can use either annotation to indicate the dilution factor in the AnalysisSuite software. If you enter that value into the “Dilution” column, the software will give you the copies/µL in your starting material (stock).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
For example, say you want to add 1 µL of a sample that has been diluted 1:10 from the stock. If you add 1 µL of your sample to a 16 µL (final volume) reaction, the dilution factor of the sample is 1:16 or 1/16 = 0.0625. Since the stock has also been diluted 1:10 (0.1), you also need to factor this in. The final dilution factor to enter into the software is 0.0625 x 0.1 = 0.00625 (1:160). As you can see below, you can use either annotation to indicate the dilution factor in the AnalysisSuite software. If you enter that value into the “Dilution” column, the software will give you the copies/µL in your starting material (stock).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.