Notification Bar Icon

Unlock 5% OFF: Shop online now and save on every purchase! Limited time offer.

Unlock 5% OFF: Shop online now and save on every purchase! Limited time offer.Learn more

Learn more
Only %1 left
SKU

NZYEasy Cloning & Expression kit IX

Storage Conditions:
-30 °C to -15 °C
NZYEasy Cloning & Expression kit IX was designed to allow directional cloning of any PCR-generated fragment or synthetic gene into the linearized pHTP9 E. coli expression vector (contains the Green Fluorescent Protein, GFP, as fusion tag).
Availability
Ready to Ship
From €29.00
NZYEasy Cloning & Expression kits were designed to allow directional cloning of any PCR-generated fragment or synthetic gene into a linearized pHTP9 Escherichia coliexpression vector. Cloning proceeds 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 pHTP vector, also containing complementary overhangs, in the presence of NZYEasy enzyme mix, 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 E. colicells. The system allows achieving high cloning efficiency (80-100%) and does not require the use of DNA ligases. In addition, the insert does not require any preliminary treatment (e.g. restriction digestion, phosphorylation, or blunt-end polishing). Cloning is performed using conventional E. colistrains. Once pHTP9 recombinant plasmid has been constructed and its sequence confirmed it should be used to transform λDE3 E. colilysogens, such as BL21(DE3), for high levels of protein expression. NZYEasy Cloning & Expression kits have 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. NZYTech provides a comprehensive portfolio of pHTP expression vectors, which include different fusion tags commonly used to enhance expression and/or solubility of recombinant proteins in E. coli, as well as fluorescent tags. pHTP1 vector (included in NZYEasy Cloning & Expression kit I, cat. No. MB281) contains two poly-histidine (6xHis) sequences (N- and C-terminal) which allow subsequent recombinant protein purification by immobilized metal ion affinity chromatography (IMAC). The other pHTP expression vectors were constructed by inserting fusion tags (see Table 1) into the pHTP1 backbone such that the fusion partner will be at the N-terminus of the recombinant protein.
More Information
Shipping Conditions Dry Ice
Storage Conditions -30 °C to -15 °C
Application Ligation Independent Cloning
Features Bacterial protein expression, Cloning, Expressed in fusion with GFP, High-throughput
Product Category DNA Cloning & Mutagenesis
Protocol time Approx. 90 min
– 10x Reaction Buffer
– NZYEasy enzyme mix
– Linearized pHTP9 vector
– Positive control
More Information
Shipping Conditions Dry Ice
Storage Conditions -30 °C to -15 °C
Application Ligation Independent Cloning
Features Bacterial protein expression, Cloning, Expressed in fusion with GFP, High-throughput
Product Category DNA Cloning & Mutagenesis
Protocol time Approx. 90 min
loader
More Information
Shipping Conditions Dry Ice
Storage Conditions -30 °C to -15 °C
Application Ligation Independent Cloning
Features Bacterial protein expression, Cloning, Expressed in fusion with GFP, High-throughput
Product Category DNA Cloning & Mutagenesis
Protocol time Approx. 90 min
More Information
Shipping Conditions Dry Ice
Storage Conditions -30 °C to -15 °C
Application Ligation Independent Cloning
Features Bacterial protein expression, Cloning, Expressed in fusion with GFP, High-throughput
Product Category DNA Cloning & Mutagenesis
Protocol time Approx. 90 min

MSDS

Material Safety Data Sheets

CoA

Certificate of Analysis

Please insert your lot number to get the available CoA