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TOPO TA for Subcloning
Quick and efficient PCR cloning with TOPO technology
TOPO TA subcloning kits utilize our popular pCR 2.1-TOPO TA vector for the direct ligation of Taq-amplified PCR products in a 5-minute, benchtop reaction yielding up to 95% recombinants.
The pCR 2.1-TOPO TA vector features:
- T7 promoter and M13 forward and reverse primer sites for in vitro RNA transcription and sequencing
- EcoRI sites flanking the PCR product insertion site for easy excision of inserts
- 15 convenient and validated restriction sites flanking your insert for easy, directional sub-cloning
- Kanamycin and ampicillin resistance for your choice of selection in E. coli
- Easy blue⁄white colony screening for selection of recombinants
- Includes One Shot TOP10 competent cells for added convenience and highest cloning efficiency
Performance and value
Each TOPO TA Cloning Kit contains linearized and topoisomerase I-activated pCR 2.1-TOPO Vector, Salt Solution, dNTPs, Control Template and Primers, M13 Forward and Reverse Primers, One Shot Chemically Competent or Electrocomp E. coli, S.O.C. Medium, and a supercoiled control plasmid. TheTOPO TA Cloning Kit for Subcloning without competent cells allows the flexibility of using your own strain for increased flexibility so competent cells and S.O.C. Medium are not provided.
In addition, TOPO TA Cloning Kits are available in combo kits combined with a PureLink Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO-cloned inserts for downstream analysis.
Using a proofreading enzyme?
Thermostable polymerases containing extensive 3´ to 5´ exonuclease activity, such as Platinum SuperFi, do not leave 3´ A-overhangs. PCR products generated with Taq polymerase have a high efficiency of cloning in the TOPO TA subcloning kits because the 3´ A-overhangs are not removed. However, if you use a proofreading polymerase or wish to clone blunt-ended fragments, you can add 3´ A-overhangs by incubating with Taq at the end of your cycling program.
Alternatively, you may want to try Zero Blunt TOPO PCR Cloning Kits. These kits offer fast and efficient cloning of blunt-end PCR products generated using thermostable, proofreading polymerases. They also utilize the ccdB gene for positive selection.
Ordering information
| Cat. No. | Product description | Competent cells supplied | Number of reactions | |
|---|---|---|---|---|
| 451641 | TOPO TA Cloning Kit for Subcloning | None | 10 | Order now |
| K4500J10 | TOPO TA Cloning Kit for Subcloning | TOP10 (genotype) | 10 | Order now |
| 450641 | TOPO TA Cloning Kit for Subcloning, without chemically competent cells | None | 25 | Order now |
| K450001 | TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Chemically Competent E. coli | TOP10 (genotype) | 25 | Order now |
| K452001 | TOPO TA Cloning Kit with One Shot MAX Efficiency DH5α T1 Phage-Resistant Chemically Competent E. coli | MAX Efficiency DH5α-T1R (genotype) | 25 | Order now |
| K455001 | TOPO TA Cloning Kit for Subcloning, with One Shot TOP10F' Chemically Competent E. coli | TOP10F' (genotype) | 25 | Order now |
| K456001 | TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Electrocomp E. coli | Electrocompetent TOP10 (genotype) | 25 | Order now |
| K451020 | TOPO TA Cloning Kit for Subcloning, with One Shot Mach1 Phage-Resistant Chemically Competent E. coli | Mach1 T1R (genotype) | 25 | Order now |
| K450002 | TOPO TA Cloning Kit with One Shot TOP10 Chemically Competent E. coli and PureLink Quick Plasmid Miniprep Kit | TOP10 (genotype) | 25 | Order now |
| K451022 | TOPO TA Cloning Kit for Subcloning, with One Shot Mach1 Phage-Resistant Chemically Competent E. coli and PureLink Quick Plasmid Miniprep Kit | Mach1 T1R (genotype) | 25 | Order now |
| K450040 | TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Chemically Competent E. coli | TOP10 (genotype) | 50 | Order now |
| K452040 | TOPO TA Cloning Kit with One Shot MAX Efficiency DH5α-T1 Phage-Resistant Chemically Competent E. coli | MAX Efficiency DH5α-T1R (genotype) | 50 | Order now |
| K455040 | TOPO TA Cloning Kit for Subcloning, with One Shot TOP10F' Chemically Competent E. coli | TOP10F' (genotype) | 50 | Order Now |
| K456040 | TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Electrocomp E. coli | Electrocompetent TOP10 (genotype) | 50 | Order now |