How to use the PCR cycle calculator
- Enter your initial DNA concentration (ng/µL). This is the amount of template DNA you start with.
- Enter your desired final concentration (ng/µL). This is the target amount of amplified DNA you want to achieve.
- Specify the reaction volume (µL) of your PCR. The default is 20 µL, but you can adjust this based on your protocol.
- Enter the PCR efficiency (percentage). A typical PCR efficiency is 90-100%. If you're unsure, use 100%.
- Click the "Calculate" button to determine the required number of PCR cycles.
Understanding PCR Cycle Calculation
The calculator uses the formula:
Cycles = log2((Desired Concentration × Reaction Volume) / (Initial Concentration × Reaction Volume)) × (1 / (1 - Efficiency/100))
This formula calculates the number of cycles needed to amplify your DNA target to the desired concentration based on the amplification efficiency.
Practical Tips
- PCR efficiency is an important parameter. A higher efficiency means more amplification per cycle, requiring fewer cycles.
- The calculator provides an estimate. It's always good to optimize your PCR conditions for the specific experiment.
- If your desired concentration is very high, you might need to consider limiting the number of cycles to avoid non-specific amplification.