NEB-style ligation setup: solve mode computes insert_ng = ratio × vector_ng × (insert bp/vector bp); check mode reports the actual pmol ratio. Recommended bands by insert size (5–10:1 under 200 bp, 3–5:1 to 1 kb, 1–5:1 above), concatemer warning above 10:1, and nM concentrations from the reaction volume. NEB / Promega / Sambrook derived. Educational use only.
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Key facts
Category
Education
Input types
select, number
Output type
json
Sample coverage
4
API ready
Yes
Overview
The Ligation Insert:Vector Molar Ratio Calculator helps plan DNA ligation reactions by calculating the insert mass needed for a target molar ratio or checking the actual ratio from vector and insert masses. It also reports pmol values, recommended ratio bands, and optional nM concentrations from the reaction volume.
When to use
Calculate the insert mass required for a chosen insert:vector molar ratio.
Check whether existing vector and insert masses fall within the recommended ratio range.
Estimate vector and insert concentrations in nM when the total reaction volume is known.
How it works
1Choose Solve insert mass to calculate the required insert mass, or Check actual ratio to compare two supplied masses.
2Enter the vector size, vector mass, insert size, and target ratio or insert mass.
3The calculator converts DNA masses and fragment sizes to pmol and determines the insert:vector molar ratio.
4Optionally enter the reaction volume to calculate vector and insert concentrations in nM.
Use cases
Planning a standard cloning ligation with a defined insert:vector ratio.
Checking whether measured DNA masses match a recommended ligation range.
Comparing DNA end concentrations in a specified ligation volume.
Examples
1. 3:1 ligation setup for a 1 kb insert
Molecular biology student
Background
A 3,000 bp vector will be ligated with a 1,000 bp insert using 100 ng of vector and a 3:1 molar ratio.
Problem
Find the insert mass required and review the resulting pmol and nM values.
How to use
Select Solve insert mass, then enter 3,000 bp vector size, 100 ng vector mass, 1,000 bp insert size, ratio 3, and a 20 µL reaction volume.