Enzymes & cloning

Ligation / assembly reaction setup

Work out how many microlitres of vector and insert to pipette to hit a target molar ratio, from each part's length and stock concentration. Handles one insert or several with independent equivalents (Gibson, Golden Gate, MoClo), reports pmol and ng per part alongside the volumes, and flags the two things that actually go wrong on a bench: a volume below what a pipette measures reliably, and a plan whose DNA does not leave room for buffer and enzyme. A molar ratio is about moles, so a shorter insert at 3 molar equivalents goes in at LESS mass than the vector — that conversion is the point.

post/ligation_setup

Request

  • Base URL: https://seqbench.com/api/v1
  • URL: https://seqbench.com/api/v1/ligation_setup
  • Auth: none required

Request body

vectorLengthBpinteger required

Backbone length in bp.

vectorNgPerUlnumber required

Backbone stock concentration, ng/µL.

vectorNgnumber

How much backbone to put in the reaction, ng. This is the quantity you fix; every insert mass follows from the ratio.

insertLengthBpinteger

Insert length in bp. Use inserts instead for a multi-fragment assembly.

insertNgPerUlnumber

Insert stock concentration, ng/µL.

molarRationumber

Molar equivalents of insert per vector. 3 is the usual starting point for a sticky-end ligation; 1-2 is common for Gibson and Golden Gate.

reactionVolumeUlnumber

Total reaction volume, µL.

vectorSequencestring

Optional backbone sequence — gives an exact molar mass from base composition instead of the 650 g/mol/bp average.

Response

Tool ran successfully.

okunknown required
toolstring required

The tool slug that ran.

resultobject required

Tool-specific output object. Its shape depends on the tool — call GET /{tool} or see the docs for each tool's fields.

Example response

{
  "provenance": {
    "apiVersion": "1.1.0"
  }
}

Changes

No recorded changes to this operation across all 1 revision of this API.