1. What is PCR Primer Concentration?

PCR primer concentration is a critical parameter in molecular biology that affects the efficiency and specificity of PCR reactions. It's typically measured in micromolar (μM) units and calculated based on the amount of primer (in nmol) and the volume of solution (in μL).

2. How Does the Calculator Work?

The calculator uses the following equation:

Where:

• \( C \) — Primer concentration (μM)
• \( nmol \) — Amount of primer (nmol)
• \( V \) — Volume of solution (μL)

Explanation: The equation converts nmol to pmol (by multiplying by 1000) and divides by the volume in μL to get concentration in μM.

3. Importance of Primer Concentration

Details: Optimal primer concentration is essential for successful PCR. Too low concentration may result in no product, while too high may cause nonspecific amplification.

4. Using the Calculator

Tips: Enter the amount of primer in nmol and the volume of solution in μL. Both values must be positive numbers.

5. Frequently Asked Questions (FAQ)

Q1: What is a typical primer concentration for PCR?
A: Most PCR reactions use primer concentrations between 0.1-1.0 μM, with 0.5 μM being common.

Q2: How do I measure the amount of primer?
A: Primer synthesis services typically provide the amount in nmol. You can also measure by spectrophotometry.

Q3: Should I use the same concentration for both primers?
A: Generally yes, unless specifically designing an asymmetric PCR.

Q4: Can I use this for other oligonucleotides?
A: Yes, this calculation works for any oligonucleotide (primers, probes, etc.).

Q5: How should I store primer stocks?
A: Store concentrated stocks at -20°C and working dilutions at 4°C for short-term use.