NOTICE: You are viewing a page of the openwetware wiki. Our "dewikify" feature makes a wiki page appear as a normal web page. In April 2017, this feature will GO AWAY and this URL will redirect to the source URL on our wiki. We're sorry for the inconvenience.

home        research        people        papers        teaching        join us        internal        pictures        contact       


ligation

ligation mix:

  • x µl digested vector
  • y µl PCR fragment or digested insert
  • 1 µl l0x ligase buffer
  • 1 µl T4 DNA ligase [3 Weiss U/µl]
  • up to 10 µl dH2O

incubate O/N at 16ºC.

in most cases, a 1:1 or a 1:3 ratio work best


formular: [(ng vector) x (kb size of insert)] / (kb size of vector) x [molar ratio of (insert/vector)] = ng insert

example: 500bp insert to be ligated with 100ng of 3.0kb vector in a 3:1 ratio
→ [(100ng vector) x (0.5 kb of insert)] / [3.0 kb vector x (3/1)] = 50 ng insert

or simply use the vector:insert ratio calculator