1. What is the Primer Reconstitution Formula?

The primer reconstitution formula calculates the volume of solvent needed to dissolve a primer to achieve a desired concentration. This is essential for molecular biology experiments where precise primer concentrations are required.

2. How Does the Calculator Work?

The calculator uses the primer reconstitution formula:

Where:

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

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

3. Importance of Proper Primer Reconstitution

Details: Accurate primer reconstitution is crucial for PCR, sequencing, and other molecular biology techniques. Incorrect concentrations can lead to failed experiments or suboptimal results.

4. Using the Calculator

Tips: Enter the amount of primer in nmol (as provided by the manufacturer) and your desired working concentration in μM. The calculator will determine how much solvent (usually TE buffer or nuclease-free water) to add.

5. Frequently Asked Questions (FAQ)

Q1: What is a typical working concentration for primers?
A: Most applications use 10-100 μM working solutions, with 100 μM being common for stock solutions.

Q2: How should I store my reconstituted primers?
A: Store at -20°C in small aliquots to avoid repeated freeze-thaw cycles.

Q3: What solvent should I use for reconstitution?
A: TE buffer (pH 8.0) or nuclease-free water are most common. Check manufacturer recommendations.

Q4: How long do reconstituted primers last?
A: Properly stored primers are typically stable for 1-2 years at -20°C.

Q5: Should I vortex primers after reconstitution?
A: Gently flick or invert the tube to mix. Avoid vigorous vortexing which can damage the DNA.