This calculator is designed for life science professionals to calculate the efficiency and molar ratios of DNA ligation reactions, which is crucial in cloning and other genetic studies.
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Data Source and Methodology
All calculations are based on standard molecular biology formulas and protocols. For more information, see NEB BioCalculator. All calculations are rigorously based on the formulas and data provided by this source.
The Formula Explained
\[ \text{Ratio} = \frac{\text{Insert Length} \times \text{Insert Conc.}}{\text{Vector Length} \times \text{Vector Conc.}} \]
Glossary of Variables
- Insert Length: The length of the DNA insert in base pairs.
- Vector Length: The length of the vector in base pairs.
- Insert Concentration: The concentration of the insert in nanograms per microliter.
- Vector Concentration: The concentration of the vector in nanograms per microliter.
How It Works: A Step-by-Step Example
For an insert of 1000 bp with a concentration of 50 ng/µL, and a vector of 3000 bp with a concentration of 20 ng/µL, the calculated insert:vector ratio is:
\[ \text{Ratio} = \frac{1000 \times 50}{3000 \times 20} = 0.83 \]
Frequently Asked Questions (FAQ)
What is DNA ligation?
DNA ligation is the process of joining two DNA strands together using a ligase enzyme.
Why is the insert:vector ratio important?
The correct insert:vector ratio is crucial for successful cloning, ensuring optimal efficiency and yield.
What concentrations should I use?
Concentrations often depend on the specific protocol and kit used, but typically range from 10-100 ng/µL.
How do I measure DNA concentration?
DNA concentration can be measured using spectrophotometry or fluorometry.
Can I use this calculator for RNA?
No, this calculator is specifically designed for DNA ligation calculations.
What if my DNA is degraded?
Degraded DNA can lead to lower ligation efficiency. Ensure DNA integrity using agarose gel electrophoresis.
Formula (LaTeX) + variables + units
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\text{Ratio} = \frac{\text{Insert Length} \times \text{Insert Conc.}}{\text{Vector Length} \times \text{Vector Conc.}}
\text{Ratio} = \frac{1000 \times 50}{3000 \times 20} = 0.83
\[ \text{Ratio} = \frac{\text{Insert Length} \times \text{Insert Conc.}}{\text{Vector Length} \times \text{Vector Conc.}} \]
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- NEB BioCalculator — nebiocalculator.neb.com · Accessed 2026-01-19
https://nebiocalculator.neb.com/
Last code update: 2026-01-19
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