1. What is IDT Reconstitution?

IDT (Integrated DNA Technologies) oligos are typically shipped as dried pellets. Reconstitution is the process of dissolving these oligos in an appropriate buffer to achieve a desired concentration for experimental use.

2. How Does the Calculator Work?

The calculator uses the IDT reconstitution formula:

Where:

• \( V \) — Volume of buffer to add (μL)
• \( OD \) — Optical density measurement (OD units)
• \( conc \) — Desired concentration (μM)
• \( dilution\_factor \) — Factor to account for any additional dilution needed

Explanation: The equation calculates the volume of buffer needed to resuspend an oligo to achieve a specific concentration, accounting for any additional dilution requirements.

3. Importance of Proper Reconstitution

Details: Accurate reconstitution is crucial for ensuring consistent experimental results. Incorrect concentrations can lead to failed experiments or misleading data in applications like PCR, sequencing, or cloning.

4. Using the Calculator

Tips: Enter the OD value from your oligo synthesis report, your desired working concentration, and any dilution factor needed. All values must be positive numbers.

5. Frequently Asked Questions (FAQ)

Q1: What buffer should I use for reconstitution?
A: TE buffer (10 mM Tris, 1 mM EDTA, pH 8.0) is commonly used, but follow your specific experimental requirements.

Q2: How should I store reconstituted oligos?
A: Store at -20°C for long-term storage. Avoid repeated freeze-thaw cycles by aliquoting.

Q3: What if my oligo doesn't dissolve completely?
A: Vortex briefly and heat at 55°C for 1-5 minutes, then vortex again. Avoid excessive heating.

Q4: How accurate is the OD measurement?
A: OD measurements are generally accurate for calculating concentration, but for critical applications, consider verifying by other methods.

Q5: Can I use this for modified oligos?
A: This calculator works for standard DNA oligos. Modified oligos may require different calculations.