Ligation Calculator
One who is struggling to find out how much DNA is needed for a ligation? and looking for easy steps on how do you determine the required insert mass? Can use our handy tool called ligation calculator. It gives output in a matter of seconds by taking the necessary inputs & also provides detailed solutions.
FAQs on Online Ligation Calculation Example with Steps
1. How do you calculate ligation using a calculator?
In order to calculate how much DNA to add to a ligation reaction at a faster pace, we have to use the ligation calculator neb. Using the ligation ne bio calculator is very simple, it asks you to enter the required inputs and hit the calculate button to obtain the desired results.
2. What is the ligation calculator formula?
The formula that our ligation calculator is using to examine the optimal mass of insert and vector needed in a cloning reaction is as follows:
Required insert mass = [(Vector mass x Insert length)/Vector length] x Molar ratio
3. What are the default required vector-to-insert molar ratios?
The Insert/vector molar ratios used for calculating the required insert mass are as such:
- 1:1,
- 2:1,
- 3:1,
- 5:1,
- 7:1
What is Ligation Reaction?
Tieing of two DNA fragments via the formation of a phosphodiester bond is known as ligation in molecular biology. The purpose of ligation is to form a recombinant plasmid.
To make it possible, we should tie off the two sticky ends covalently which are the DNA backbone and the DNA insert. It is catalyzed by an enzyme, known as a ligase that catalyzes the ligation reaction. Also, the important part of DNA replication is Ligation.
Tips to Optimize DNA Ligation Reactions for Cloning
There are a few basic and useful tips that enhance the ligations and also may improve the cloning success rate. Check out the below tips & try to implement them:
1. Lower the temperature
2. Check the pH
3. Aliquot the Ligase Buffer
4. Include a Restriction Enzyme Just Before Transformation
5. Heat the DNA Just Before Ligation
6. Add PEG (Polyethylene Glycol)
Steps of the DNA Ligation Reaction
The following are the two important & basic steps that itself DN ligation reaction own:
- Initially, the ends of DNA need to collide by chance and coalesce for much time for ligase to unite.
- The next step is to perform the enzymatic reaction.
Ligation Calculator Formula
The formula to calculate the ligation molar ratio considering the vector mass and length, insert length, and the ratio of insert/vector is given by
Required insert mass = (Vector mass * Insert length)/Vector length * Molar ratio
Where units for the above formula,
- Micrograms (μg) or Nanograms (ng) for the required insert mass & given Vector mass
- Base pairs(bp) or Kilobases (kb) for the Insert length & vector length
- The molar ratio is for insert/vector ratio
If you want to know what is the formula used to calculate the DNA Concentration then make sure to check out our DNA Concentration Calculator & also use all math calculator tools from onlinecalculator.guide for easy calculations.
How to Use the Ligation Calculator?
The simple steps to make you understand how to calculate the ligation ratio with our free online ligation calculator are here:
- Visit the ligation calculator and enter the input values like insert length, vector length, vector mass, and insert/vector molar ratio.
- Also, select the units of the input with the help of the dropdown list available.
- Do pick any of the insert/vector molar ratios from the provided list or else use the ideal ratio which is 3:1.
- After entering all inputs & their units select the unit for the desired result and then click on the calculate button to display the output in no time.