5-minute cloning with positive selection
TOPO TA Cloning kits for sequencing utlize our popular pCR™4-TOPO TA Vector for the direct ligation of Taq-amplified PCR products in a 5-minute, benchtop reaction yielding up to 95% recombinants.
pCR™4-TOPO allows you to directly select recombinants by disrupting the lethal E. coli gene, ccdB. The vector contains the ccdB gene fused to the C-terminus of the LacZα fragment. Ligating a PCR product disrupts expression of the lacZα-ccdB gene fusion permitting growth of only positive recombinants upon transformation in competent cells that contain non-recombinant vector are killed upon plating. Therefore, blue/white screening is not required.
The pCR™4-TOPO TA Vector—Optimized for Sequencing
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Performance and value
Each TOPO TA Cloning Kit for sequencing contains linearized and topoisomerase I-activated pCR™4-TOPO TA 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. The TOPO TA Cloning Kit for sequencing 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, the TOPO TA Cloning Kit for Sequencing, with One Shot TOP10 Chemically Competent E. coli, and PureLink™ Quick Plasmid Miniprep Kit allows for 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 Pfx, 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.
订购信息
| 目录号 | 产品描述 | Competent cells supplied | 反应次数 | |
|---|---|---|---|---|
| K4575J10 | TOPO TA Cloning Kit for Sequencing, with One Shot TOP10 Chemically Competent E. coli | TOP10 (genotype) | 10 | 立即订购 |
| 450030 | TOPO TA Cloning Kit for Sequencing, without competent cells | 无 | 20 | 立即订购 |
| K457501 | TOPO TA Cloning Kit for Sequencing, with One Shot TOP10 Chemically Competent E. coli | TOP10 (genotype) | 20 | 立即订购 |
| K458001 | TOPO TA Cloning Kit for Sequencing, with One Shot TOP10 Electrocomp™ E. coli | Electrocompetent TOP10 (genotype) | 20 | 立即订购 |
| K459501 | TOPO TA Cloning Kit for Sequencing, with One Shot MAX Efficiency DH5α T1 Phage-Resistant Chemically Competent E. coli | MAX Efficiency DH5α (genotype) | 20 | 立即订购 |
| K453020 | TOPO TA Cloning Kit for Sequencing, with One Shot Mach1™ T1 Phage-Resistant Chemically Competent E. coli | Mach1™ T1R (genotype) | 20 | 立即订购 |
| K457502 | TOPO TA Cloning Kit for Sequencing, with One Shot TOP10 Chemically Competent E. coli, and PureLink™ Quick Plasmid Miniprep Kit | TOP10 (genotype) | 20 | 立即订购 |
| K457540 | TOPO TA Cloning Kit for Sequencing, with One Shot TOP10 Chemically Competent E. coli | TOP10 (genotype) | 40 | 立即订购 |
| K458040 | TOPO TA Cloning Kit for Subcloning, with One Shot TOP10 Chemically Competent E. coli | Electrocompetent TOP10 (genotype) | 40 | 立即订购 |
| K459540 | TOPO TA Cloning Kit for Sequencing, with One Shot MAX Efficiency DH5α T1 Phage-Resistant Chemically Competent E. coli | MAX Efficiency DH5α (genotype) | 40 | 立即订购 |
仅供科研使用，不可用于诊断目的。