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Technology

Azura Genomics is primarily engaged in the following scientific areas:



End-Point PCR
Invented in 1983 by Kary Mullis, end-point polymerase chain reaction (PCR) enables researchers to expand a single piece of DNA into millions of identical fragments. This well-established technique makes it possible to detect sequences that might otherwise be undetectable. In recent years, the molecular biologist has asked progressively more challenging questions and PCR applications and complexity have evolved. Azura Genomics has developed end-point PCR systems which deliver best-in-class sensitivity, speed and yield. Azura scientists have identified improvements in enzyme solubility and novel buffer additives which enhance reaction efficiency and processivity.

Clearly, our reagents are different and deliver superior performance in PCR applications such as genotyping, colony PCR, high-fidelity PCR, crude sample PCR, multiplex PCR, GC-rich PCR, and long PCR. For example, Azura 2x HS Taq Mix is a robust, single-tube Hot-Start formulation which has been fully optimized for a broad range of applications and capable of fast cycling and room temperature reaction assembly for convenience and throughput. Our Azura TruFi™ DNA Polymerase is a new generation, ultra High-Fidelity DNA Polymerase which provides greater yields with lower enzyme amounts than other polymerases, even in crude PCR reactions with known inhibitors. TruFi™ is ideal for high-fidelity PCR for cloning, mutagenesis, next-generation re-sequencing, and long PCR up to 10kb.


Gene Expression and qPCR
Today, real-time PCR is considered the gold standard for accurate, sensitive and fast measurement of gene expression. Real-Time PCR, also referred to as quantitative PCR (qPCR) was developed as a highly sensitive, rapid and efficient method for nucleic acid detection. This ever-popular technique is based on conventional PCR with the following improvements:

  • qPCR combines amplification and detection in a single step.
  • It has an increased limit of detection and therefore can use smaller amounts of starting material than traditional PCR.
  • The DNA produced can be quantified based on fluorescent detection.

Azura Genomics has engineered real-time PCR solutions which deliver unrivalled sensitivity with low-copy gene targets and robust performance in the presence of common PCR inhibitors. In addition, our scientists have developed a unique buffer chemistry including novel additives for advanced multiplexing capability, delivering the same high efficiency, early Ct and reaction speed in complex multiplex qPCR assays as standard single-plex reactions. The proprietary buffer systems with our unique balance of salts, pH and stabilizers allow for exceptional performance with GC-rich and AT-rich sequences and the option of ultra-fast cycling. One of our core technologies is high-temperature reverse transcription and the generation of accurate relative cDNA representation regardless of gene abundance. The AzuraQuant™ cDNA Synthesis Kit provides complete, unbiased cDNA representation from as little as 4pg Total RNA for subsequent Real-Time PCR and the convenience of a simple, two-component kit. For workflows which involve One-Step RT-qPCR, Azura Genomics has developed the ideal reaction chemistry and cycling parameters for efficient first-strand cDNA synthesis and subsequent real-time PCR in the same reaction mixture, providing a seamless and convenient method for high-throughput gene expression studies. For example, the AzuraQuant™ Probe One-Step qPCR Kit NoRox can be used to quantify low abundance RNA targets while reducing the number of pipetting steps and time to result. It is ideally suited to pathogen detection, multiplex RT-qPCR, viral quantitation, and biomarker discovery.



Genotyping
Genetic variation is present in many different forms in the genome, including somatic mutations, single nucleotide polymorphisms (SNPs), copy number variations (CNVs) and structural changes. In order to study and better understand the many variations, Azura Genomics has developed an offering of powerful genotyping tools. High resolution Melt (HRM) and dual-probe based analysis are common methods used in SNP genotyping. We have applied recent advances in enzyme preparation in conjunction with a careful consideration of ionic conditions and buffer components to deliver clear allele discrimination for higher confidence genotyping. The AzuraQuant™ HRM Fast Mix contains Azura HS Taq polymerase, high affinity antibodies and Vivid-Green™ 2 dye, a novel saturating, third-generation dye which is ideal for accurate SNP genotyping. It has been developed and optimized for fast PCR cycling, improved sensitivity, and maximum discrimination between sequence variants.

Alternatively, the use of transgenic mice has long been a cornerstone for genetic experimentation. Transgenic mice are commonly used in the lab to model human diseases such as diabetes and cancer. The many challenges of purifying high quality, intact DNA from transgenic mice and subsequently amplifying target genes have been addressed with limited success. Azura Genomics has developed a simple, cost-effective direct PCR platform for high-throughput mouse genotyping. Our polymerase chemistry is ideally suited to high-performance with crude samples, and a broad range of amplicon lengths and sequence complexity. The Azura Mouse Genotyping Kit includes a proprietary lysis buffer formulation which delivers a rapid and complete release of genomic DNA from mouse tail snips or ear punches in 15 minutes. This is followed by PCR amplification using Azura 2x HS Red Mix for unrivalled sensitivity and performance regardless of GC-rich and AT-rich DNA regions. Azura 2x HS Red Mix also includes an inert red dye which facilitates direct gel-loading.



Cloning
Successful cloning and subsequent protein expression requires highly efficient DNA transformation. The process of altering a cell so that it acquires and expresses genetic material from a surrounding environment is transformation. This process is commonly used to introduce recombinant plasmid DNA into bacterial strains which can be made competent by artificial means. Chemically competent cells are calcium chloride-treated to facilitate attachment of the plasmid DNA to the competent cell membrane. Cells are subsequently heat-shocked which opens the pores in the cell membrane so plasmid DNA from the surrounding environment can enter the cell. Electrocompetent cells are prepared for transformation using electroporation for a few milliseconds. This method uses an electrical pulse to create pores in the membrane so that the material can enter the cell from the surrounding environment.

Azura Genomics has screened an exhaustive list of compounds and additives which strongly influence the permeability of the cell membrane in chemical transformation and electroporation. Our High5™ Alpha Chemically Competent E. coli represent an advance in providing consistent, superior transformation efficiency cells for the most demanding cloning applications such as the construction of gene banks or cDNA libraries with limited amounts of DNA. In comparison to other commercially available competent cells, High5™ Alpha Chemically Competent E. coli and High5™ Alpha Electrocompetent E. coli provide significantly higher transformation efficiencies and convenient packaging for ease-of-use.

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