Target CloneTM and Target CloneTM -Plus- are TA cloning kits developed by Toyobo for rapid, efficient, and authentic cloning of PCR products.
This manual describes the TA cloning system developed by Toyobo for rapid and reliable insertion of PCR products into the pTA2 vector. It covers the A‑attachment Mix reaction, ligation setup, transformation of competent cells, and verification of recombinant colonies.
Detailed protocols are given for PCR product preparation, optional A‑addition, ligation conditions, competent‑cell handling, heat‑shock transformation, and colony PCR screening. Safety notes and storage recommendations for reagents are also included. This manual is written in Chinese.
| Minimum ligation time | 5 minutes |
|---|---|
| Maximum insert size | 12 kb |
| A‑attachment Mix reaction time | 10 minutes at 60 °C |
| pTA2 vector concentration | 50 ng/µl |
The ligation reaction can be completed in as little as 5 minutes.
Up to 12 kb PCR products can be cloned using this system.
Store the pTA2 vector at –20 °C and avoid long periods at room temperature or repeated freeze‑thaw cycles.
Incubate the reaction at 60 °C for 10 minutes.