Quick and efficient PCR cloning with TOPO® TA cloning

Clone PCR-amplified DNA fragments (blunt or A-overhang) directly into a choice of over 40 subcloning, sequencing, or expression vectors in just 5 minutes—and obtain up to 95% recombinant clones.

  • Quick—avoid inefficient ligation and laborious searches for appropriate restriction enzymes
  • Efficient—minimize colony screening by obtaining up to 95% recombinant clones
  • Troubleshooting tool—if it can’t be cloned with TOPO® cloning technology, it can't be cloned

 

TOPO® cloning protocol


Figure 1. The TOPO® cloning protocol. TOPO® PCR cloning requires just three easy steps. Simply combine your PCR product and a TOPO® cloning vector in the provided solution, wait five minutes, then transform E. coli.

Steps TOPO® TA Cloning Kits TA/UA Cloning Restriction enzyme or cut-back cloning
Use of existing primers? Yes Yes No
Vector is ready for cloning? Yes Yes No
Ligation regents included? Yes Yes No
Prepare or purchase competent cells separately? No, included Purchase: 0 hr
Prepare: up to 6 hr
Purchase: 0 hr
Prepare: up to 6 hr
Ligation time 5 min 1 hr 2 - 23 hr
Recombination efficiency Up to 95% 60 - 80% ~60%
Time required for cloning 5 min 1 - 12 hr 2 - 23 hr