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TOPO TA Cloning
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
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
| 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 reagents 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 |