Molecular biology

Ligation ratio

Insert mass for a target insert:vector molar ratio.

ng insert = ratio x ng vector x (insert bp / vector bp)

Vector mass (ng)

Vector length (bp)

Insert length (bp)

Insert : vector ratio

Protocol result

⚗ Aliquot

Ligation ratio · insertNg = molarRatio · vectorNg · (insertBp / vectorBp) · the lab bench, calculated

Ligation✓ insertNg = molarRatio · vectorNg · (insertBp / vectorBp)

Calculated live

Use 30 ng of insert with 50 ng of vector (3:1).

Combine 30 ng insert, 50 ng vector, ligase and buffer.

Vector

50 ng

Ratio

3:1

Insert needed

30 ng