Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms

Journal Article · Sat Apr 01 00:00:00 EST 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.86.8.2766· OSTI ID:5229255

Orita, Masato; Iwahana, Hiroyuki; Kanazawa, Hiroshi; Hayashi, Kenshi; Sekiya, Takao ^[1]

---

1. National Cancer Center Research Institute, Tokyo (Japan)

The authors developed mobility shift analysis of single-stranded DNAs on neutral polyacrylamide gel electrophoresis to detect DNA polymorphisms. This method follows digestion of genomic DNA with restriction endonucleases, denaturation in alkaline solution, and electrophoresis on a neutral polyacrylamide gel. After transfer to a nylon membrane, the mobility shift due to a nucleotide substitution of a single-stranded DNA fragment could be detected by hybridization with a nick-translated DNA fragment or more clearly with RNA copies synthesized on each strand of the DNA fragment as probes. As the mobility shift caused by nucleotide substitutions might be due to a conformational change of single-stranded DNAs, we designate the features of single-stranded DNAs as single-strand conformation polymorphisms (SSCPs). Like restriction fragment length polymorphisms (RFLPs), SSCPs were found to be allelic variants of true Mendelian traits, and therefore they should be useful genetic markers. Moreover, SSCP analysis has the advantage over RFLP analysis that it can detect DNA polymorphisms and point mutations at a variety of positions in DNA fragments. Since DNA polymorphisms have been estimated to occur every few hundred nucleotides in the human genome, SSCPs may provide many genetic markers.

• OSTI ID: 5229255
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 86:8, Issue 8; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DNA
ELECTROPHORESIS
RFLPS
DNA HYBRIDIZATION
BLADDER
CARCINOMAS
DNA SEQUENCING
GENE MUTATIONS
LEUKOCYTES
MAN
MELANOMAS
PHOSPHORUS 32
PLASMIDS
TUMOR CELLS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY
BODY FLUIDS
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
DISEASES
HYBRIDIZATION
ISOTOPES
LIGHT NUCLEI
MAMMALS
MATERIALS
MUTATIONS
NEOPLASMS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS ISOTOPES
PRIMATES
RADIOISOTOPES
STRUCTURAL CHEMICAL ANALYSIS
URINARY TRACT
VERTEBRATES
550200* - Biochemistry