1. What is Primer Resuspension?

Primer resuspension is the process of dissolving lyophilized oligonucleotides (primers) in an appropriate buffer to achieve a desired working concentration, typically in micromolar (μM) units.

2. How Does the Calculator Work?

The calculator uses the primer resuspension formula:

Where:

• \( C \) — Primer concentration (μM)
• \( nmol \) — Amount of primer in nanomoles
• \( V \) — Resuspension volume in microliters

Explanation: The formula converts the primer amount from nmol to pmol (by multiplying by 1000) and divides by the volume to get concentration in μM.

3. Importance of Primer Concentration

Details: Accurate primer concentration is crucial for PCR efficiency, ensuring proper annealing temperatures and preventing non-specific amplification.

4. Using the Calculator

Tips: Enter the amount of primer (as provided by the manufacturer) in nmol and your desired resuspension volume in μL. Both values must be positive numbers.

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: What buffer should I use for resuspension?
A: TE buffer (10 mM Tris, 1 mM EDTA, pH 8.0) or nuclease-free water are commonly used.

Q3: How should I store resuspended primers?
A: Store at -20°C for long-term storage. Avoid repeated freeze-thaw cycles by aliquoting.

Q4: How do I dilute my primer to a working concentration?
A: Use the formula: Volume of stock = (desired concentration × desired volume) / stock concentration.

Q5: How can I verify my primer concentration?
A: UV spectrophotometry (A260 measurement) can be used to verify concentration after resuspension.