1. What is Fusion Molar Ratio?

The Fusion Molar Ratio represents the ratio of moles of DNA insert to moles of vector DNA in a ligation reaction. It's crucial for efficient cloning, with typical ratios ranging from 2:1 to 10:1 (insert:vector).

2. How Does the Calculator Work?

The calculator uses the following formula:

Where:

• moles of insert = (insert mass in ng × 10⁻⁹) / (insert length in bp × 660 g/mol)
• moles of vector = (vector mass in ng × 10⁻⁹) / (vector length in bp × 660 g/mol)
• 660 g/mol — Average molecular weight of a DNA base pair

Explanation: The calculation converts mass to moles using the molecular weight of DNA and length of each fragment.

3. Importance of Molar Ratio

Details: Proper molar ratios are essential for efficient ligation. Too much insert can lead to multiple inserts, while too little can result in empty vectors.

4. Using the Calculator

Tips: Enter the mass (in ng) and length (in bp) for both insert and vector. Typical insert:vector ratios range from 2:1 to 10:1 for most cloning applications.

5. Frequently Asked Questions (FAQ)

Q1: What is the ideal molar ratio for cloning?
A: For most applications, a 3:1 to 5:1 insert:vector ratio works well, but this may vary depending on insert and vector characteristics.

Q2: Why is 660 g/mol used for DNA calculations?
A: This is the average molecular weight of a DNA base pair (sodium salt), accounting for variations between AT and GC base pairs.

Q3: Should I use ng or µg for input?
A: The calculator is designed for nanograms (ng), which is the typical unit for small-scale DNA measurements.

Q4: How does insert length affect the ratio?
A: Longer inserts have more mass per mole, so you need more mass of a short insert than a long one to achieve the same molar ratio.

Q5: Can I use this for RNA or ssDNA?
A: No, this calculator is for double-stranded DNA. RNA and single-stranded DNA have different molecular weights per base.