High-Throughput Gene Cloning

NZYTech offers a high-throughput cloning service for large projects, typically involving several hundreds to thousands of genes. A proprietary method is used for cloning your genes into one or more selected vectors. Our method does not require the use of conventional restriction enzymes and DNA ligases, which increases speed while improving cloning efficiencies, as only the desired DNA fragments are cloned into the destination vector(s) in the correct orientation.

A DNA template source provided by the customer will be used for gene amplification, by PCR. Alternatively, genes can be chemically synthesized (at an additional cost). DNA fragments are cloned into standard cloning vectors, and this cloning method allows to easily exchange genes from an entry vector (pHTP0) into various pHTP Escherichia coli expression plasmids (see figure below). NZYTech has an own library of several pHTP expression plasmids containing different tags, known to enhance protein’s solubility and/or promote correct protein’s folding. This strategy may constitute the best alternative for selecting the most appropriate tag for each protein.

^{PCR fragments or synthetic genes cloned into an entry vector can be easily transferred into any pHTP E. coli expression vector. The NZYTech’s portfolio of pHTP expression vectors includes a different range of fusion tags, thus offering the possibility to quickly assay levels of expression and solubility of a desired protein in multiple conditions}.
Features: - Genes of interest are obtained from gDNA, cDNA, pDNA, phage, cosmid DNA or, alternatively, by gene synthesis - Fast cloning procedure - QC by verification of correct plasmids sequence and frame using Sanger sequencing - Flexibility: gene sub-cloning can be easily achieved from the pHTP0 cloning vector to any of the pHTP derivatives with different fusion tags - High protein expression levels can be reached when using pHTP expression vectors (see expression levels) - Considerable saving of time and money - Customers retain all rights to the sequence data and related intellectual property - Absolute confidentiality is guaranteed
Delivery Specifications: - 0.5-1 μg of purified recombinant plasmids containing your genes (in 96-well plates) - HTP Gene Cloning Report: includes QC, vector map and insert DNA sequences - Estimated delivery: upon request