▶ EZchange™ Site-directed Mutagenesis kit primer design program

Product description

EZchange™ Site-directed Mutagenesis kit allows site- specific mutations with virtually any double-stranded plasmid as template, and thus eliminates a time-consuming subcloning of target DNA into single stranded DNA-producing plasmids. In addition, no specialized vectors or unique restriction sites are required. Therefore, mutations can be produced in a simple and convenient way.

Characteristics

- No subcloning

- High efficiency

- Site-direted mutagenesis in a single day

Applications

- Substitution

- Single or multiple deletions

- Insertion mutation

Kit contents

- nPfu-Frote

- 10X Reaction Buffer

- dNTP mixture (2 mM each)

- EZ-MIX

- 2X EZ-MIX Buffer

- Control primers (10 pmol/μl)

- Control Plasmid (5ng/μl)

- Sterile water

Standard reaction conditions

- Mutant Strand Synthesis Reaction

^aTemplate plasmids should be purified from dam⁺ E. coli strains. Mutation efficiency would decrease markedly if plasmids isolated from dam^- strains are used, since they are not digested by Dpn l. Most E. coli strains used in laboratories are dam⁺ except JM110 and SCS110.

Cautions: Adjust the temperature of the reaction tube to 94℃ prior to the addition of nPfu-Forte. The high temperature prevents nonspecific amplification or unwanted primer degradation. As a result, the mutation efficiency is significantly increased.

- EZ-MIX reactions

→ Incubate the reaction mixture for 30 min at 37℃

- Transformation

 Use 1-2 μl of DNA treated with EZ-MIX for transformation and spread 1/100 to 1/10 of transformed  

cells on appropriate plates.

●  Enzynomics_EZ004_EZchange™ Site-directed Mutagenesis Kit_manual_kor

●  Enzynomics_EZ004_EZchange™ Site-directed Mutagenesis Kit_manual_simple(eng)

● Material Safety Data Sheet

- Enzynomics_MSDS_EZ004_EZchange™ Site-directed Mutagenesis Kit_E 

● Related paper

1. S-M Ahn, et al(2012). Oncogene 31, 3051-3059

2. Hayeong Kwon, et al(2009). Biochim Biophys Acta 1793, 1325-1333

3. Huong T. T. Ngo, et al(2013). J. Virol. 87, 5718-5731

4. Hoyun Choi, et al(2013). J Virol. 15, 03159-12

5. Hayeong Kwon, et al(2010). J Cell Mol Med 15, 888-908