TOPO® TA Cloning
Quick and efficient PCR Cloning with TOPO®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.
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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 Cloning® Kits deliver the highest efficiency in the shortest time.
| 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 |
NEW! TOPO® Kits available without competent cells
Select the best TOPO® Cloning Kit for your research
Whether you’re doing general subcloning, sequencing, in vitro transcription, or expression in E.coli, mammalian cells, or our Gateway® system, there’s a TOPO® cloning solution for you.
- General Subcloning: Convenient, validated restriction sites in the pCR® 2.1 vector simplify future sub-cloning
- Sequencing: Minimal MCS region in our pCR® 4 vector ensures you sequence more insert , less vector the first time
- In vitro transcription: Both SP6 and T7 both promoters in pCR® II vector makes IVT reactions convenient
- Mammalian expression: pcDNA™ TOPO® vectors are the industry standard for constitutive expression
- TOPO® Cloning of Long PCR Fragments: Combining TOPO®, ZeroBackground™ and Crystal Violet technologies make cloning easy for 3-10 kb fragments
- TOPO® Cloning into Gateway® Entry Vectors: Direct access to our suite of Gateway® expression vectors
