1. What is Infusion Molar Ratio?

The Infusion Molar Ratio calculation determines the appropriate amount of DNA insert needed relative to the vector DNA for cloning experiments. This ensures optimal ligation efficiency during molecular cloning procedures.

2. How Does the Calculator Work?

The calculator uses the molar ratio formula:

Where:

• \( \text{vector} \) — Vector DNA amount in nanograms
• \( \text{ratio} \) — Desired molar ratio of insert to vector
• \( \text{insert length} \) — Length of insert DNA in base pairs
• \( \text{vector length} \) — Length of vector DNA in base pairs

Explanation: The equation accounts for the molecular weight difference between insert and vector DNA to calculate equivalent molar amounts.

3. Importance of Molar Ratio Calculation

Details: Proper molar ratios (typically 3:1 to 5:1 insert:vector) are critical for efficient ligation and successful cloning. Incorrect ratios can lead to poor cloning efficiency or vector self-ligation.

4. Using the Calculator

Tips: Enter vector amount in ng, desired molar ratio, and lengths of both insert and vector in base pairs. All values must be positive numbers.

5. Frequently Asked Questions (FAQ)

Q1: What is a typical molar ratio for cloning?
A: Most cloning protocols recommend a 3:1 insert:vector ratio, though some applications may require different ratios (2:1 to 5:1).

Q2: How does DNA length affect the calculation?
A: Longer DNA molecules have more mass per mole, so the calculator adjusts for length differences between insert and vector.

Q3: Should I use linear or circular vector length?
A: Use the length of the linearized vector (after restriction digest), not the circular plasmid length.

Q4: What if my insert is much smaller than the vector?
A: The calculator automatically accounts for size differences - smaller inserts require less mass to achieve the same molar ratio.

Q5: Can I use this for Gibson Assembly?
A: Yes, though Gibson Assembly often uses different ratios (typically 2:1 insert:vector for each fragment).