Description

The ligation reaction is an essential step in genetic recombination experiments. For this reaction, T4 DNA ligase has been widely used. Ligation high Ver.2 is a highly efficient premixed T4 Ligase reagent.

Features

  • Effective ligation of cohesive, blunt and A overhang DNA fragments can be achieved.
  • Will not freeze at -20ºC. No need to thaw.
  • Just mix ligation high Ver.2 with an equal volume or with double the volume of the solution containing DNA fragments.

Applications

  • Ligation of DNA frangments

Storage condition

Store at -20ºC*

*In the case of -30ºC storage or long-term storage under -20ºC, the reagent might freeze. No decrease in reaction efficiency is observed following
5 freeze-thaw cycles.

Components

Ligation high Ver.2 750µl (100 reactions)

Application data

Example 1.TA cloning using purified PCR products

T vector,(50 ng, 25 fmol,) was mixed, at different ratios, with purified PCR products (500 bp from Taq DNA polymerase). An equal volume of Ligation high Ver.2, 7.5 µl, was added and incubated at 16ºC for 30min. E. coli DH5α competent cells were transformed using 10 µl of reaction mixture, and cultured on LB/Amp(X-Gal, IPTG) plates O/N.

Fig. 1.TA cloning using purified products

Colony number at 1:5 corresponds to 100

Example 2. TA Cloning using unpurified PCR products

T vector,(50 ng, 25 fmol,) was mixed, at different ratios, with unpurified PCR products (500 bp from Taq DNA polymerase). An equal volume of Ligation high Ver.2, 7.5 µl, was added and incubated at 16ºC for 30 min. E. coli DH5α competent cells were transformed using 10 µl of reaction mixture, and cultured on LB/Amp(X-Gal, IPTG) plates O/N.

Fig. 1. TA cloning using unpurified products

Colony number on the condition of adding 0.5µl of the PCR product corresponds to 100.