# Ligation Insert:Vector Molar Ratio Calculator 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. > Canonical page: https://elysiatools.com/en/tools/ligation-molar-ratio - **Category:** Education - **Keywords:** ligation molar ratio, insert to vector ratio calculator, ligation calculator neb, dna ligation setup, insert mass ng ligation, vector insert molarity, pmol dna ligation, t4 dna ligase ratio, cloning insert calculator, dephosphorylated vector ligation, molecular biology, lab calculator ## 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. ## Inputs - **Calculation Mode** (select): Solve for the insert mass a desired ratio needs, or check the actual ratio of two masses. - **Vector Size (bp)** (number): Double-digested vector backbone length in base pairs. - **Vector Mass (ng)** (number): Mass of the linearized vector in the reaction. - **Insert Size (bp)** (number): Insert fragment length in base pairs. - **Insert Mass (ng)** (number): Insert mass (check mode); ignored in solve mode, which computes the required value. - **Desired Insert:Vector Ratio** (number): Molar insert:vector ratio (solve mode), e.g. 3 for a 3:1 setup. - **Reaction Volume (µL, optional)** (number): Total ligation volume in µL; enter 0 to skip nM concentrations. - **Decimal Places** (number) ## 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 - Choose Solve insert mass to calculate the required insert mass, or Check actual ratio to compare two supplied masses. - Enter the vector size, vector mass, insert size, and target ratio or insert mass. - The calculator converts DNA masses and fragment sizes to pmol and determines the insert:vector molar ratio. - Optionally 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. ## Frequently asked questions ### What formula does the calculator use? Solve mode uses insert mass = target ratio × vector mass × (insert size ÷ vector size). ### What does Check actual ratio do? It uses the supplied vector and insert masses and reports their actual insert:vector molar ratio. ### What ratio is recommended for a 750 bp insert? The recommended band is 3–5:1 for inserts from 200 to 999 bp. ### When are nM concentrations calculated? Enter a reaction volume greater than 0 µL. Enter 0 to skip concentration calculations. ### What does a ratio above 10:1 indicate? 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