Manual

Product Number: CLN1102

Shipping and Storage

-Storage at 20 ℃, transportation at ≤ 0 ℃.

Component

Note: a. The positive control control contains a 4.6 kb linearized vector and a 1 kb insertion fragment with Amp resistance. When using, directly add 10μL of Control to 10μL of 2×fast Multi One-Step Cloning kit, Form a 20μL recombinant reaction system.

Description

The 2× fast Multi CloneMix is a one-step, multi fragment universal seamless cloning kit developed based on the principle of homologous recombination. This series of recombinant cloning kits belongs to a non ligase dependent system and has a very low background for carrier self ligation. The highly optimized 2× fast Multi CloneMix pre mixes enhanced recombinase and the required buffer for recombination reaction, which can significantly improve the recombination efficiency and tolerance to impurities of cloning. Therefore, the prepared high-purity linearized vector and insertion fragment can be directly used for recombinant Kron without purification, greatly simplifying the experimental operation.

This kit can selectively recombine insertion fragments containing overlapping regions at the end of the vector to any site of any vector, without being limited by enzyme cleavage sites (except in special cases such as cloning of toxic genes or instability of the plasmid after insertion of exogenous DNA). It can insert a single fragment or up to 5 sequentially spliced fragments at once. The vector end, insertion fragment end, and adjacent insertion fragment ends require a completely identical sequence of 15-25bp that can be homologous recombined with each other. Under the action of recombinase, the insertion fragment and linearized vector can be recombined by reacting at 37℃ for 5-30 minutes. The recombinant product does not rely on ligase and can be directly transformed into competent cells, with a positive cloning rate of over 95%.

Features

1. Fast: recombination can be completed within 5-30 minutes;
2. High throughput cloning: not limited by enzyme cleavage sites, no need for fragment cleavage;
3. Seamless cloning: does not introduce additional sequences.

Scope of application

Suitable for experiments such as rapid seamless cloning, site directed mutagenesis, and high-throughput cloning of 1-5 fragments.