DNA Ligation Calculator
How to Use:
1. Enter the length (bp) and mass (ng) of your Vector DNA.
2. Enter the length (bp) of your Insert DNA.
3. Set the desired Molar Ratio (Insert : Vector). (A 3:1 ratio is generally recommended).
4. Click Calculate to get the required Insert mass and a standard 20 µL reaction setup.
1. Enter the length (bp) and mass (ng) of your Vector DNA.
2. Enter the length (bp) of your Insert DNA.
3. Set the desired Molar Ratio (Insert : Vector). (A 3:1 ratio is generally recommended).
4. Click Calculate to get the required Insert mass and a standard 20 µL reaction setup.
:
Calculation Result & Reaction Setup
Required Insert DNA Mass
0.00 ng
| Component | Volume / Amount |
|---|---|
| 10X T4 DNA Ligase Buffer | 2.0 µL |
| Vector DNA ( ng) | Variable |
| Insert DNA ( ng) | Variable |
| T4 DNA Ligase | 1.0 µL |
| Nuclease-free Water | up to 20.0 µL |
| Total Reaction Volume | 20.0 µL |
* Note: Ligase buffers contain ATP, which degrades upon multiple freeze-thaw cycles. Keep aliquots small. Thaw buffer at room temperature and vortex vigorously before use.