Detection of DNA sequence polymorphisms in human genomic DNA by using denaturing gradient gel blots
- Tufts University School of Medicine, Boston, MA (United States)
Denaturing gradient gel electrophoresis can detect sequence differences outside restriction-enzyme recognition sites. DNA sequence polymorphisms can be detected as restriction-fragment melting polymorphisms (RFMPs) in genomic DNA by using blots made from denaturing gradient gels. In contrast to the use of Southern blots to find sequence differences, denaturing gradient gel blots can detect differences almost anywhere, not just at 4-6-bp restriction-enzyme recognition sites. Human genomic DNA was digested with one of several randomly selected 4-bp recognition-site restriction enzymes, electrophoresed in denaturing gradient gels, and transferred to nylon membranes. The blots were hydridized with radioactive probes prepared from the factor VIII, type II collagen, insulin receptor, [beta][sub 2]-adrenergic receptor, and 21-hydroxylase genes; in unrelated individuals, several RFM's were found in fragments from every locus tested. No restriction map or sequence information was used to detect RFMP's.
- OSTI ID:
- 7176880
- Journal Information:
- American Journal of Human Genetics; (United States), Journal Name: American Journal of Human Genetics; (United States) Vol. 50:2; ISSN AJHGAG; ISSN 0002-9297
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
COLLAGEN
CYTOLOGICAL TECHNIQUES
DNA HYBRIDIZATION
DNA SEQUENCING
ELECTROPHORESIS
ENZYMES
GENES
HORMONES
HYBRIDIZATION
HYDROXYLASES
INSULIN
MAMMALS
MAN
MEMBRANE PROTEINS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PEPTIDE HORMONES
PRIMATES
PROTEINS
RECEPTORS
RFLPS
SCLEROPROTEINS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES