Ligation Insert:Vector Molar Ratio
Insert mass to add for a target insert:vector molar ratio (insert ng = ratio x insert bp/vector bp x vector ng).
Example
You enter
- Vector mass (ng) 50
- Vector length (bp) 5000
- Insert length (bp) 1000
- Insert:vector molar ratio 3
You get
- Insert mass to add 30 ng of insert
- Amounts (pmol) 0.01538
- Insert pmol 0.04615
Checked against published worked examples from: Standard molecular cloning (ligation insert:vector molar ratio).
Details, formula, and sources
ScopeInsert mass to add for a target insert:vector molar ratio (insert ng = ratio x insert bp/vector bp x vector ng), plus each in picomoles at 650 g/mol per bp. A short insert needs less mass, not equal mass; equal masses over-represent small fragments.
Formula or tableinsert_ng = ratio x (insert_length / vector_length) x vector_ng; vector_pmol = vector_ng / (vector_length x 650) x 1e3; insert_pmol = ratio x vector_pmol.
Source and editionLigation insert:vector molar-ratio setup (standard molecular cloning), by name.
Free accessStandard molecular-cloning bench relation; the enzyme protocol and fragment ends govern.
What governsEstimate. AHJ and licensed professional govern.
API field namesvector_ng, vector_length_bp, insert_length_bp, molar_ratio, insert_ng, vector_pmol, insert_pmol
- Average bp mass 650 g/mol per base pair (dsDNA average; ssDNA and RNA differ)standard molecular cloning