1. What is the Ligation Calculator?

The Ligation Calculator Insilico Pro helps determine the optimal amount of insert DNA needed for a ligation reaction based on vector mass, lengths, and desired molar ratio.

2. How Does the Calculator Work?

The calculator uses the ligation formula:

Where:

• \( MI \) — Insert mass (ng)
• \( MV \) — Vector mass (ng)
• \( LI \) — Insert length (bp)
• \( LV \) — Vector length (bp)
• \( R \) — Molar ratio (insert:vector)

Explanation: The equation calculates the mass of insert DNA needed to achieve a specific molar ratio with the vector DNA in the ligation reaction.

3. Importance of Molar Ratio Calculation

Details: Proper molar ratios are crucial for efficient ligation. Typical ratios range from 1:1 to 10:1 (insert:vector) depending on the application.

4. Using the Calculator

Tips: Enter vector mass in ng, lengths in bp, and desired molar ratio. All values must be positive numbers.

5. Frequently Asked Questions (FAQ)

Q1: What's a typical molar ratio for ligation?
A: Common ratios range from 3:1 to 5:1 (insert:vector) for standard cloning, but may vary based on application.

Q2: How do I measure vector mass accurately?
A: Use a spectrophotometer (Nanodrop) or fluorometer (Qubit) for precise DNA quantification.

Q3: Does this work for different vector types?
A: Yes, the calculator works for any vector type (plasmids, fosmids, BACs) as long as you know the lengths.

Q4: What if I have multiple inserts?
A: Calculate each insert separately, then combine them in the desired ratios.

Q5: How does vector linearization affect this?
A: The calculator assumes both vector and insert are linear. For circular vectors, use the total length.