Experimental parameters

The traditional classical molecular cloning technology based on restriction endonuclease and T4 DNA ligase is prone to false positive and low cloning efficiency. It is also limited by many practical factors, such as the selection of enzyme digestion sites, cumbersome preparation of insertion fragments, preference of T4 DNA ligase sequence, long ligation reaction time and so on.

Seamless cloning technology does not rely on T4 ligase and is not limited by the restriction of enzyme digestion sites of vector and target fragment. The overlapping fragment recombination method is adopted. The special enzyme combination can directionally recombine and connect the vector linearized by any method with the PCR fragment with 15-25bp overlapping region at both ends, so as to quickly realize the efficient and seamless cloning of one or more fragments.

The Magic Multi s one step cloning kit adopts the form of one tube mix. It only needs one-step recombination method to obtain a highly efficient cloning recombinant vector, which greatly improves the cloning efficiency.

Product features.

1. One or more PCR amplified fragments (flat ended) clones can be inserted into any position of any vector within 30 minutes.

2. It can be cloned at any site without being limited by the availability of vector and insertion fragment enzyme digestion sites and flat end / sticky end.

3. Seamless cloning, the insertion point will not introduce unwanted base sequences.

4. Independent of ligase and phosphatase, it is efficient and accurate, and the cloning positive rate can reach more than 95%.

5. The ligation of fragments was completed by incubation at room temperature

After the transformation of single fragment and multi fragment recombinant products, more monoclonal antibodies can be produced on the plate, and almost no clones can be produced on the negative control plate without inserted fragments

20 monoclonal antibodies were selected from single fragment transformation plate for colony PCR, and the clone positive rate was more than 95%

15 monoclonal clones were selected from 4 Fragment recombinant transformation plate for colony PCR, and the clone positive rate was more than 85%

Please click the link below to download the manual

M501 seamless cloning reagent - Magic Multi s one step cloning kit pdf