NZYEasy Cloning kit

NZYEasy Cloning kit was designed to allow directional cloning of any PCR-generated fragment into a linearized pHTP0 vector in a single ligase-independent reaction mediated by the NZYEasy enzyme mix. Vector-complementary overhangs containing a specific sequence recognized by the NZYEasy enzyme are incorporated in the PCR product by using primers with appropriate 5’ extensions. When you combine the insert thus generated with the linearized pHTP0 vector, also containing complementary overhangs, in the presence of NZYEasy enzyme, the two DNA molecules will anneal through base-pair complementation of the single-strand regions. The reaction occurs in a single-tube along three temperature-dependent steps. Circular recombinant vector containing the fragment of interest is obtained by transforming the annealed plasmid DNA into competent Escherichia coli cells.

The system allows achieving high cloning efficiency (80-100%) and does not require the use of DNA ligases. In addition, no further treatment (e.g. restriction digestion, phosphorylation, or blunt-end polishing) of the inserts is required. This kit has been successfully used in high-throughput (HTP) platforms for the efficient cloning of a large number of genes at a scale compatible with the functional screen of hundreds to thousands of genes.

This kit has been successfully used in high-throughput (HTP) platforms for the efficient cloning and expression of a large number of genes at a scale compatible with the functional screen of hundreds to thousands of genes/proteins.

Features:

• Ligase-independent cloning
• Time-saving and cost-effective method
• No additional treatment of the PCR fragments required (e.g restriction digestion, phosphorylation or blunt-end polishing)
• High cloning efficiencies
• Directional cloning
• Possibility to clone multiple inserts simultaneously into one vector
• Compatibility with automation and high-throughput methods
• Suitable for both cloning or bacterial protein expression

Applications:

• Cloning of blunt-end or A-overhangs PCR products

Components:

• 10x Reaction Buffer
• NZYEasy enzyme mix
• Linearized pHTP vector
• Positive control

Manual for Multiple Fragment Cloning using pHTP vectors     

NZYEasy Cloning Expression kits Product Brochure
 
FAQ NZYEasy clonning and Expression

pHTP0 Expression Vector NZYTech