Description:
Choo-Choo Cloning^TM Kits are highly efficient directional PCR cloning kits for rapid, ligase and restriction enzyme-independent cloning of PCR products. They allow to clone any PCR fragment into any linearized vector at any location. By simply incubating on ice, the ends of a PCR-generated DNA fragment can be precisely fused into a DNA vector with 6 bp (or more) overlapping homologous sequence. The system is very robust. Up to 8 PCR-generated DNA fragments can be assembled and cloned into one piece (up to 10 kb in one step). The system is highly efficient, with 98-100% positive colonies.

Function:
The function of Choo-Choo Cloning^TM Kits depend on MCLAB's proprietary enzyme systems. There is no need for restriction enzyme digestion, ligation, or blunt-end polishing. Any extra unwanted bases can be eliminated in the final construct. The linearized vector can be generated by PCR or restriction enzyme digestion. The PCR fragments can be generated by Taq DNA polymerase or other high fidelity DNA polymerases. The addition of a A by Taq DNA polymerase is not required and has no effect on cloning efficiency. If the PCR product is amplified from a plasmid template, then it needs to be gel-purified to reduce background. In addition to PCR cloning, Choo-Choo Cloning^TM Kits also have the following aplications: adaptor, linker and tag addition before and after the inserts, and gene synthesis.

Features:
- Clone any insert into any vector you choose at any location
- No restriction digestion, phosphatase treatment, or ligation required
- Multiple fragments (up to 8 pieces)
- Broad PCR sizes (up to 10 kb)
- Final constructs are seamless with no extra or unwanted base pairs
- Simple 45 minute single-tube reaction on ice protocol
- High efficiency with >=98% positive colonies
- Multiple functions: compatible with adaptor, linker and tag addition before or after the inserts; compatible with gene synthesis
- High throughput application

Recommended Storage Conditions:
Store Box 1 at -20°C.
Store Box 2 at -80°C.

Components:
The Choo-Choo Cloning^TM Kits are available in 10-, 20-, 100-, and 96-reaction sizes.