Microsatellite Genotyping
AGRF uses state-of-the-art technologies and high throughput processes for microsatellite genotyping. This service has been applied to a wide variety of species and can accommodate any species for which microsatellite markers are available. Microsatellites, due to their highly informative nature, provide a powerful tool to any linkage mapping or population genetics study. Microsatellite Genotyping Applications include:
Microsatellite Genotyping: Sample Submission Requirements
Human Genome-Wide SNP
The AGRF currently offers the medium density 10cM AB PRISM Human Linkage Mapping set for genome-wide microsatellite scans. This linkage mapping set is comprised of 400 highly polymorphic dinucleotide microsatellite markers, spaced at an average distance of 10cM throughout the human genome. Each marker is individually amplified using fluorescently labeled PCR primer pairs and then pooled into marker panels for capillary separation on the AB3730 DNA Analyser. Genotyping results are analyzed using AB GeneMapper software.
Human Genome-Wide SNP: Sample Submission Requirements
Human Genome-Wide Scan Service Process
Fine Mapping
For fine mapping studies the AGRF can provide technical expertise to assist the researcher in selecting informative markers relevant to the appropriate area of interest on the human genome. The AGRF is able to assist in the selection of microsatellite markers from the high density 5cM and medium density 10cM AB human linkage mapping sets. In cases where the markers in either of these linkage mapping sets don't suit the specific chromosomal location or density requirements, the AGRF offers a custom genotyping service for fine mapping. The AGRF has experience and expertise to provide assistance in the selection of:
- microsatellites,
- primer design and ordering
- paneling of custom microsatellite markers.
Additionally, a mixture of AB human linkage mapping set and custom microsatellite markers can be selected to achieve the requirements specific to your fine mapping projects needs.
Fine Mapping Service Process
Custom Genotyping
AGRF has extensive experience in providing support for the selection and design of microsatellite markers for a wide range of plant and animal species. AGRF offers a full service which includes assistance in designing and ordering fluorescently labeled primer pairs for the microsatellite markers of your choice. In cases where multiple markers are to be genotyped, this can be done in a cost-effective manner whereby AGRF can panel markers for capillary separation. Genotyping results are analysed using AB GeneMapper software for the calling of either standard or customised alleles.
Custom Genotyping Service Process
Fragment Analysis
The fragment analysis service offered by the AGRF provides both capillary separation and analysis services for fluorescently labeled DNA fragments. An AB3730 DNA analyser is used for capillary separation and data is analyzed using AB GeneMapper software. Marker types applied to fragment analysis include microsatellites, AFLP (amplified fragment length polymorphisms) and T-RFLP (terminal restriction fragment length polymorphisms), for which analysis can be customised to your marker type and project requirements.
For this service the customer performs PCR and provides the AGRF with PCR product for electrophoresis. The AB3730 DNA Analyser platform can accommodate up to four different coloured fluorescent dyes: 6-FAM, VIC, NED or PET. This allows multiple markers to be amplified (for the same individual), pooled and then run in a single capillary. Where multiple markers of the same colour are pooled, it is important to ensure that the fragment size range of markers does not overlap. A maximum fragment length of 1,000 base pairs can be analyzed on the AB3730 DNA Analyser.
Fragment Analysis Service Process
