Topo TA克隆方法(Topo TA Cloning Kit) Topo TA克隆原理与TA克隆一样,唯一不同的是TA克隆用的是T4连接酶把PCR片断连接到T载体上,而Topo TA Cloning用的是DNA Topoisomerase。 Topoisomerase的用途一般使用在复制DNA前把超螺旋DNA切割使之解旋后,再连接成线性DNA。 Topo TA克隆即使用Topoisomerase高效连接的特性把含3`A端的PCR扩增片断快速连接到3`T端载体上,整个反应只需五分钟!一般T4连接酶需过夜才能高效连接。Topo TA克隆系统提供Topoisomerase I载体,感受态细胞,SOC培养基等。 Topo平端克隆方法和长PCR片断克隆法(Zero Blunt Topo PCR Cloning Kit)(Topo XL PCR Cloning Kit) |
TOPO PCR表达克隆法
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表1 PCR克隆试剂盒的选择 |
Amplification Enzyme | Application of Cloned PCR Product | Ligation Method | Product | Advantages | Cat. No. |
Taq DNA Polymerase | •Sequencing | TOPO® Cloning (5 minutes) Topoisomerase | TOPO TA Cloning® Kit | •≥95% recombinants | K4500-01 |
TOPO TA Cloning® Kit Dual Promoter | •≥95% recombinants | K4600-01 | |||
TOPO TA Cloning® Kit for Sequencing | •≥95% recombinants | K4575-01 | |||
Ligase-mediated(overnight)T4 DNA ligase | Original TA Cloning® Kit | •≥80% recombinants | K2000-01 | ||
Original TA Cloning® Kit Dual Promoter | •≥80% recombinants | K2050-01 | |||
TOPO® Cloning(5 minutes) Topoisomerase | Various | •Fast cloning directly into an expression vector | All the TOPO Exp. Kits | ||
Proofreading polymerase | •Expression of cloned PCR product | TOPO® Cloning(5 minutes) Topoisomerase | Zero Blunt® TOPO® PCR Cloning Kit | •≥95% recombinants | K2800-20 |
Zero Blunt® TOPO® PCR Cloning Kit for Sequencing | •≥95% recombinants | K2875-20 | |||
Ligase-mediated(1 hour)T4 DNA ligase | Zero Blunt® PCR Cloning Kit | •≥95% recombinants | K2700-20 | ||
Polymerase mixturesfor long Template混合酶 | •Sequencing | TOPO® Cloning(5 minutes) Topoisomerase | TOPO® XL PCR Cloning Kit | •High-efficiency cloning of long (3-10kb) PCR products | K4700-10 |