Molecular biology

How do you calculate gibson / ligation planner?

Vector amount and insert ratios to an exact pipetting table. The relationship used is V = (vector_pmol · ratio) / (stock_ng/µL · pmol_per_ng).

Worked example

Every number below is computed by the calculator, not typed in

What you are given

  • Vector amount (pmol): 0.05
  • Reaction volume (µL): 20
  • Fragments: [{"name":"Vector","role":"vector","length":"5000","stock":"50","ratio":"1"},{"name":"Insert","role":"insert","length":"1000","stock":"25","ratio":"2"}]

Formula: V = (vector_pmol · ratio) / (stock_ng/µL · pmol_per_ng)

The working

  1. Vector: target = 0.05 pmol
  2. Per fragment: pmol = vector · ratio; V = pmol / (ng/µL · 1000/(bp·660))

Pipette 2 fragments into 20 µL.

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