GenieClone DNA Assembly Site-Directed Mutagenesis Kit v1

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SKU:
MORV0005
€117

Description

ELISA Kit Technical Manual

Product Information

GenieClone Site-Directed Mutagenesis Kit v1 is designed for rapid site-directed mutagenesis of up to 2 separate mutations in the same reaction in less than 3 hours. Based on GenieClone rapid cloning technology, this kit uses homologous recombination to replace the conventional annealing ring-forming reactions meaning much less template and a more flexible primer design strategy. With seamless splicing of the two PCR products by GenieClone technology, the kit can complete up to two separate mutations with a single amplification reaction. DpnI digested products of the specific amplicons can be directly added to the recombination reaction without purification. The highly optimized reaction buffer, fast protocol and superior site-directed mutagenesis efficiency make GenieClone Site-Directed Mutagenesis Kit v1 make an excellent choice for introducing 1 or 2 mutations at discontinuous sites simultaneously on the same plasmid.

 
 

Figure 1: Single base site-directed mutation using GenieClone Site-Directed Mutagenesis Kit v1

Key Features

  • Introduce up to 2 separate mutations in the same reaction.
  • Site-directed mutagenesis can be performed on up to 2 discontinuous sites (more than 50 bp apart) simultaneously on the target plasmid.
  • Single kit for cloning and mutagenesis.
  • Includes Genie Fusion Ultra High-Fidelity DNA Polymerase providing high-fidelity PCR with extremely low mutation rates & excellent long fragment amplification for any plasmid amplification up to 20 kb as well as GC-rich templates.
  • Amplification is carried out exponentially and the template usage is extremely low, which is beneficial to the complete degradation of the original methylation template.
  • DpnI eliminates contamination of the original template and decreases background.
  • Amplified products can be directly used in the recombination reaction after DpnI treatment.
  • The reaction mixture can be used to directly transform chemically competent E.coli cells.

Figure 2: Schematic of introducing single base or continuous multi-bases site-directed mutation into plasmid

Pack Information

Components MORV0005 - 10 rxn MORV0005 - 25 rxn

2 x Max Buffer

1.25 ml

1.25 ml

dNTP Mix (10 mM each)

20 µl

50 µl

Genie Fusion Ultra High-Fidelity DNA Polymerase

20 µl

50 µl

Dpnl (10 U/µl)

20 µl

50 µl

5 x CE II Buffer

40 µl

100 µl

GenieClone Recombinase

20 µl

50 µl

Note: use methylase non-defective host strains (eg. Top10, DH5α, JM109) to extract the original plasmids for use with this kit.

Storage

Store the product at -20°C, and it will be valid for 1 year.

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