1. What is IDT Primer Resuspension?

IDT (Integrated DNA Technologies) primer resuspension involves dissolving lyophilized primers in an appropriate volume of buffer or water to achieve the desired concentration for molecular biology applications.

2. How Does the Calculator Work?

The calculator uses the standard resuspension formula:

Where:

• \( nmol \) — Amount of primer in nanomoles
• \( 10 \) — Standard conversion factor (10 µl per nmol)

Explanation: This formula provides a standard 100 µM concentration when resuspending primers, which is the typical working concentration for most molecular biology applications.

3. Importance of Proper Resuspension

Details: Accurate resuspension is crucial for achieving consistent primer concentrations, ensuring reproducible results in PCR, sequencing, and other molecular techniques.

4. Using the Calculator

Tips: Enter the nmol value provided on the IDT primer tube. The calculator will determine the appropriate volume of resuspension buffer or water needed to achieve standard concentration.

5. Frequently Asked Questions (FAQ)

Q1: Why use 10 µl per nmol?
A: This standard ratio produces a 100 µM stock solution, which is the recommended concentration for most molecular biology applications.

Q2: What type of solution should I use for resuspension?
A: Use nuclease-free water or TE buffer (10 mM Tris, 1 mM EDTA, pH 8.0) for optimal primer stability and performance.

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

Q4: Can I use different concentrations?
A: While 100 µM is standard, you can adjust the volume to achieve different concentrations based on your specific experimental needs.

Q5: How do I verify primer concentration after resuspension?
A: Use a spectrophotometer to measure absorbance at 260 nm and calculate the exact concentration using the primer's extinction coefficient.