High-Throughput Gene Cloning

Storage Conditions:

-85 °C to -15 °C

For orders above 24 clones, NZYtech specializes in large projects, offering a comprehensive library of E. coli expression plasmids. These vectors feature diverse tags, optimizing protein solubility and folding for an economical and flexible solution.

Availability

Produced On Demand

Details

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

More Information
Shipping Conditions	Dry Ice
Storage Conditions	-85 °C to -15 °C

Figures & Tables

High-throughput Gene Synthesis Pipeline

Codon optimization of your genes

Proteins play a variety of critical roles in many biological processes. They are valuable therapeutic compounds, research reagents and industrial catalysts. NZYtech's proprietary codon optimization software – ATGenium - can alter both naturally occurring and recombinant gene sequences to achieve the highest possible levels of protein expression in any given expression system. This will save you time and money, while getting you quickly to meaningful results. ATGenium takes into consideration a variety of critical factors involved in different stages of protein expression, such as codon adaptability, mRNA structure, and various cis-elements in transcription and translation. Optimizing your gene sequence will have significant advantages in a variety of applications such as recombinant protein expression, DNA vaccine design and protein engineering.

Optimizing heterologous protein production in Escherichia coli with ATGenium:
Superior levels of recombinant proteins expressed in Escherichia coli strains can be achieved through codon optimization using the NZYtech's ATGenium algorithm.

A. Levels of protein expression of three different eukaryotic proteins in a prokaryotic system (Escherichia coli). The genes encoding these proteins were optimized by three algorithms for codons optimization, by two different competitors (light blue) and using the ATGenium algorithm developed by NZYtech (dark blue).

B. High-throughput recombinant protein expression of CAZYmes (Carbohydrate Active eNZymes). Genes encoding these proteins were optimized using ATGenium algorithm and overexpressed in Escherichia coli BL21 (DE3). Recombinant proteins were purified through IMAC and separated by SDS-PAGE. High protein expression yields were obtained when genes were optimized using ATGenium algorithm.