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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.
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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 presenceof 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.
Shipping Conditions | Dry Ice |
---|---|
Storage Conditions | -30 °C to -15 °C |
Application | Ligation Independent Cloning |
Features | Bacterial protein expression, Cloning, High-throughput |
Product Category | DNA Cloning & Mutagenesis |
Protocol time | Approx. 90 min |
– 10x Reaction Buffer
– NZYEasy enzyme mix
– Linearized pHTP0 vector
– Positive control
– NZYEasy enzyme mix
– Linearized pHTP0 vector
– Positive control
Shipping Conditions | Dry Ice |
---|---|
Storage Conditions | -30 °C to -15 °C |
Application | Ligation Independent Cloning |
Features | Bacterial protein expression, Cloning, High-throughput |
Product Category | DNA Cloning & Mutagenesis |
Protocol time | Approx. 90 min |
https://www.nzytech.com/wp-content/uploads/2016/04/NZYEasy-Cloning-Method.jpg
Figure 1. Schematic representation of the cloning procedure following NZYEasy Cloning kit. After amplification with primers that include overhangs to append to the gene-specific sequence, PCR insert is treated with NZYEasy enzyme mix to promote annealing of the insert to the linearized pHTP vector containing complementary ends.
https://www.nzytech.com/wp-content/uploads/2016/04/Multiple_cloning.jpg
Figure 2. Illustration of multiple fragment cloning using the NZYEasy Cloning & Expression System. The figure exemplifies the cloning of three PCR-generated inserts (fragments 1, 2 and 3) into a pHTP vector using NZYEasy enzyme mix. Complementary sequences required to correctly assemble the full length nucleic acid are highlighted in light green, dark blue, magenta and dark green.
Figure 1. Schematic representation of the cloning procedure following NZYEasy Cloning kit. After amplification with primers that include overhangs to append to the gene-specific sequence, PCR insert is treated with NZYEasy enzyme mix to promote annealing of the insert to the linearized pHTP vector containing complementary ends.
https://www.nzytech.com/wp-content/uploads/2016/04/Multiple_cloning.jpg
Figure 2. Illustration of multiple fragment cloning using the NZYEasy Cloning & Expression System. The figure exemplifies the cloning of three PCR-generated inserts (fragments 1, 2 and 3) into a pHTP vector using NZYEasy enzyme mix. Complementary sequences required to correctly assemble the full length nucleic acid are highlighted in light green, dark blue, magenta and dark green.
Shipping Conditions | Dry Ice |
---|---|
Storage Conditions | -30 °C to -15 °C |
Application | Ligation Independent Cloning |
Features | Bacterial protein expression, Cloning, High-throughput |
Product Category | DNA Cloning & Mutagenesis |
Protocol time | Approx. 90 min |
Shipping Conditions | Dry Ice |
---|---|
Storage Conditions | -30 °C to -15 °C |
Application | Ligation Independent Cloning |
Features | Bacterial protein expression, Cloning, High-throughput |
Product Category | DNA Cloning & Mutagenesis |
Protocol time | Approx. 90 min |
Shipping Conditions | Dry Ice |
---|---|
Storage Conditions | -30 °C to -15 °C |
Application | Ligation Independent Cloning |
Features | Bacterial protein expression, Cloning, High-throughput |
Product Category | DNA Cloning & Mutagenesis |
Protocol time | Approx. 90 min |
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