Clone PCR-amplified DNA fragments

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 Invitrogen 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.

TOPO TA compared to traditional cloning methods

StepsTOPO TA Cloning KitsTA/UA CloningRestriction enzyme or cut-back cloning
Use of existing primers?YesYesNo
Vector is ready for cloning?YesYesNo
Ligation reagents included?YesYesNo
Prepare or purchase competent cells separately?No, includedPurchase: 0 hr
Prepare: up to 6 hr
Purchase: 0 hr
Prepare: up to 6 hr
Ligation time5 min1 hr2–23 hr
Recombination efficiencyUp to 95%60–80%~60%
Time required for cloning5 min1–12 hr2–23 hr