A first generation factory for typing DNA polymorphisms
- Marshfield Medical Research Foundation, Marshfield, WI (United States); and others
Although linkage mapping of fully penetrant, single-gene disorders can readily be accomplished using {open_quotes}manual{close_quotes} polymorphism genotyping methods, mapping of genes responsible for genetically more complex disorders such as asthma and schizophrenia will likely require more highly automated approaches. Routine clinical application of linkage analysis will also require reliable automated methods. To meet these needs we have initiated a first generation factory for analysis of short tandem repeat polymorphisms. The factory includes an eight-pronged pipetting robot for efficient PCR setup, amplification within high density microtiter plates, and a scanning fluorescence detection system for automatic sizing of amplification products. Initial throughput for the factory will be several hundred thousand genotypes per year. The heart of the factory is a new scanning fluorescence detection system for gel electrophoresis. The system employs an argon laser for excitation and has the capability of simultaneously detecting four different fluorescent dyes. The detector scans across the bottom of short, wide denaturing polyacrylamide gels which routinely contain 144 lanes. Emitted fluorescent light is collected with a high numerical aperture microscope objective and transmitted through a fiber optic cable, dichroic mirrors and band pass filters to a series of photomultiplier tubes. Amplified output from the photomultiplier tubes is digitized and fed into computers for further analysis. Software has been written for viewing the gel images, for lane finding and for allele identification. Rhodamine-labeled {lambda} DNA standard fragments ranging in size from 70 to 300 bases are utilized for lane identification and allele sizing. Each gel run takes 3 hours, and each gel can be loaded at least three times. Multiple markers can be amplified simultaneously and detected without prior concentration.
- OSTI ID:
- 134136
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-0872
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
BASIC STUDIES
99 MATHEMATICS
COMPUTERS
INFORMATION SCIENCE
MANAGEMENT
LAW
MISCELLANEOUS
44 INSTRUMENTATION
INCLUDING NUCLEAR AND PARTICLE DETECTORS
HEREDITARY DISEASES
GENES
HUMAN CHROMOSOMES
GENETIC MAPPING
DNA
MUTATIONS
STRUCTURAL CHEMICAL ANALYSIS
DETECTION
AUTOMATION
BIOLOGICAL MARKERS
GENOTYPE
POLYMERASE CHAIN REACTION
FLUORESCENCE
ELECTROPHORESIS
COMPUTER CODES