Ligation Calculator — vector : insert setup

V_insert = R × (C_vector × V_vector) × L_insert ÷ (C_insert × L_vector) · length in bp stands in for molar mass

1 · DNA stocks

Vector concentration ng/µL

Insert concentration ng/µL

Vector length bp

Insert length bp

2 · Molar ratio — vector : insert

1 : 1 1 : 2 1 : 3 1 : 5

Custom ratio R For 1 : 3, enter 3

3 · What to hold fixed

Vector volume Vector mass

Vector volume to pipette µL Usually 1–2 µL

4 · Reaction volumeoptional

Total reaction µL

Enzyme + buffer µL From your kit's manual

Set the total to 0 to skip the water calculation.

Pipetting scheme 1 : 3

Vector—
—µL
Insert—
—µL
Enzyme + bufferas specified
—µL
Nuclease-free waterto final volume
—µL
DNA subtotalvector + insert
—µL

Volumes are workable Both volumes clear 0.5 µL.