1. What is Primer Reconstitution?

Primer reconstitution is the process of dissolving lyophilized primers in an appropriate volume of buffer or water to achieve a desired concentration for molecular biology experiments.

2. How Does the Calculator Work?

The calculator uses the primer reconstitution formula:

Where:

• \( V \) — Volume of solvent needed (μL)
• \( nmol \) — Amount of primer (nmol)
• \( C \) — Desired concentration (μM)

Explanation: The formula converts nmol to pmol (×1000) and divides by the desired concentration in μM to get the volume in μL needed to dissolve the primer.

3. Importance of Proper Primer Reconstitution

Details: Accurate primer reconstitution is essential for consistent PCR results, proper primer concentrations in reactions, and reproducible experimental outcomes.

4. Using the Calculator

Tips: Enter the amount of primer in nmol (typically provided by the manufacturer) and your desired working concentration in μM. Both values must be positive numbers.

5. Frequently Asked Questions (FAQ)

Q1: What is a typical working concentration for primers?
A: Most PCR applications use primers at 10-20 μM, but this depends on your specific protocol.

Q2: Should I use TE buffer or water for reconstitution?
A: TE buffer (pH 8.0) helps maintain primer stability, but nuclease-free water is acceptable for short-term use.

Q3: How should I store reconstituted primers?
A: Store at -20°C in aliquots to avoid repeated freeze-thaw cycles. For long-term storage, keep lyophilized primers at -20°C or -80°C.

Q4: What if my primer amount is in μg instead of nmol?
A: You'll need to convert using the molecular weight of the primer. The formula is: nmol = (μg × 1000) / (MW × number of bases).

Q5: How accurate does the volume need to be?
A: For most applications, ±5% is acceptable. Use an accurate pipette and ensure complete dissolution by vortexing.