Newpep FastClone kit is a fast cloning kit that does not require enzyme cutting or ligation. It can clone PCR products into any position of the vector without being restricted by the enzyme cutting site. The kit can clone up to 20 kb of DNA fragments into the vector with a positive rate of more than 90%, which greatly improves the existing cloning efficiency.
The FastClone Kit uses homologous recombination to insert PCR products into any position of a vector without restriction enzymes or T4 DNA ligase. It enables cloning of DNA fragments ranging from 100 bp to 20 kb, achieving positive rates of over 90 % and reducing reaction time to about 30 minutes.
The kit includes FastClone Enzyme, 10X FastClone Buffer, EcoRI‑linearized pUC19 plasmid, and a 1 kb positive‑control PCR product. Detailed protocols cover vector linearization, preparation of insert fragments with 15 bp vector‑matching overhangs, and directional subcloning with a recommended 1:5 vector‑to‑insert molar ratio. This manual is written in Chinese.
| Reaction time | 30 minutes |
|---|---|
| Positive cloning rate | up to 90 % |
The cloning reaction is completed in about 30 minutes at room temperature.