Molecular biology

Gibson / ligation planner

Vector amount and insert ratios to an exact pipetting table.

V = (vector_pmol · ratio) / (stock_ng/µL · pmol_per_ng)
NEBuilder suggests ~0.03 to 0.06 pmol of vector.
FragmentRoleLength (bp)Stock (ng/µL)Ratio
Protocol result
⚗ Aliquot
Gibson / ligation planner · V = (vector_pmol · ratio) / (stock_ng/µL · pmol_per_ng) · the lab bench, calculated
Assembly✓ V = (vector_pmol · ratio) / (stock_ng/µL · pmol_per_ng)
Calculated live

Pipette 2 fragments into 20 µL.

1

Vector: 3.3 µL (0.05 pmol).

2

Insert: 2.64 µL (0.1 pmol).

FragmentpmolVolume (µL)
Vector (vector)0.053.3
Insert (insert)0.12.64
Water14.06
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Gibson / ligation planner, Aliquot