Systems
Transgenomic WAVE® Systems utilize DHPLC (denaturing high-performance liquid chromatography) in conjunction with the Transgenomic patented separation DNASep® Cartridges for mutation detection. WAVE Systems offer discovery and detection of genetic variation at close to 100% sensitivity— making it one of the most sensitive and accurate technologies for detection of known and unknown mutations and single nucleotide polymorphisms (SNPs).
WAVE Systems are used throughout the world to screen for a large variety of diseases. More than 350 human genes have been screened entirely or partly by DHPLC. A multitude of other applications are being used with WAVE Systems in such diverse areas as plant genomics, microbial analysis and drug sensitivity. With close to 1,300 installations around the world, the WAVE System is an internationally accepted technology.
With a wide variety of models, fully automated WAVE Systems provide a robust, versatile and cost effective option, meeting the critical requirements of leading academic and biopharmaceutical laboratories.
The WAVE System allows you to discover and detect mutations and size nucleic acid fragments through denaturing high performance liquid chromatography (DHPLC). The WAVE System is more accurate and sensitive than other methods for mutation detection.
DHPLC Process
Wild and test or mutant type alleles are amplified separately, mixed, heated and then cooled to form homoduplexes and heteroduplexes (if a mutation is present).
When a mutation is present, there are four possible configurations:
The sample containing homoduplexes and heteroduplexes (if a mutation is present) is injected into the buffer flow path containing triethyammonium acetate (TEAA) and acetonitrile (ACN). In solution, the TEAA forms the positively charged triethylammonium ion (TEA+) that has both hydrophobic and hydrophilic ends. The DNASep cartridge is located in the oven and contains beads that are hydrophobic. The positively charged portion of theTEA+ forms an association with the negatively charged phosphate backbone of the DNA creating a hydrophobic "fur" on the fragment. This entire hydrophobic entity then behaves as would a typical hydrophobic molecule and is attracted to the hydrophobic beads.
The fragment specific methods created by Navigator™ Software control both the temperature of the oven and the ACN gradient. The concentration of ACN increases over time based on this method. As the ACN concentration increases bridging capabilities of the TEA+ ions decrease and the DNA fragments are released from the cartridge. Heteroduplexes, with mismatched base pairs, elute off of the cartridge first followed by the homoduplexes. The fragments pass through the UV detector which detects the absorbance over time. This information is sent to Navigator Software on the computer. It records the results in the form of a chromatogram. If no mutation is present, all of the homoduplex DNA fragments elute off of the cartridge at the same time producing a single peak on the chromatogram. If a mutation is present then two to four peaks will be visible as illustrated below.
At non-denaturing temperatures, the smaller fragments elute off of the cartridge before larger fragments allowing for fragment separation based on size.
WAVE Systems can be used for a wide range of applications including:
- Mutation Detection
- Sizing
- Oligonucleotide Purification and Analysis
- Forensics
- Microbial Analysis
- RNA Isolation and Purification
- Single Base Extension
- Methylation
With the WAVE System you can:
- Scan fragments for both known and novel mutations/SNPs without extensive resequencing
- Detect low-abundance mutations in heterogeneous samples
- Enrich for low-abundance alleles
- Fractionate complex mixtures of related fragments based on differences in sequence content
- Perform targeted mutation/SNP scoring via primer extension assays
- Analyze end-point products from allele-specific PCR, RT-PCR assays.
Systems