Cold Fusion Cloning Kit without Competent Cells

Fast, easy, and efficient - The Cold Fusion Cloning Kit will take you to transformation-ready DNA in 20 minutes with a one tube reaction format

Product Description

• Get transformation-ready DNA in as little as twenty minutes
• A phosphatase-free and ligation-free system that eliminates the need for specific restriction enzymes
• Typically achieve >90% positive clones
• Add any insert into any site in any vector using manual or automated workflows

The Cold Fusion technology is a simple, rapid and highly efficient cloning kit. It allows you to directly clone any PCR product(s) to any linearized expression vector, at any site.

Four-step Cold Fusion Process

1. Linearize your vector and amplify your insert using primers with at least 15 bp of homology to the ends of your linearized vector
2. Mix your PCR-amplified inserts with the linearized vector
3. Incubate for 5 minutes at room temperature and then 10 minutes on ice to fuse the insert to the linearized vector
4. Transform competent cells with the fusion mix. A typical Cold Fusion Cloning reaction delivers 90% positive clones

The PCR fragments can be generated by High Fidelity DNA Polymerases or Taq DNA Polymerase, with primers that are designed to have at least 15 bases of homology at their linear ends to where you want the product to fuse. The linearized vector can be generated by PCR or restriction enzymes (single or double cut). The kit is so robust that multiple DNA fragments can be assembled simultaneously and cloned into one construct in a single step. The system is highly efficient, with more than 90% positive cloning rate.

The Cold Fusion Cloning Kit is also available with competent cells and in 96-well format.

Performance Data

Efficient Cold Fusion cloning of single DNA Fragments

 Figure 1. Cold Fusion Cloning of 3′, 5′, and blunt-end fragments.

Cold Fusion cloning of multiple DNA fragments - Capable of cloning multiple fragments in the order you choose

Figure 2. Using Cold Fusion technology, the team at SBI built a 4-in-1 construct to generate iPSCs in only three days.