Alu polymerase chain reaction: A method for rapid isolation of human-specific sequences from complex DNA sources

Journal Article · Fri Sep 01 00:00:00 EDT 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.86.17.6686· OSTI ID:6968905

Nelson, D L; Ledbetter, S A; Corbo, L; Victoria, M F; Ramirez-Solis, R; Webster, T D; Ledbetter, D H; Caskey, C T ^[1]

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1. Baylor College of Medicine, Houston, TX (USA)

Current efforts to map the human genome are focused on individual chromosomes or smaller regions and frequently rely on the use of somatic cell hybrids. The authors report the application of the polymerase chain reaction to direct amplification of human DNA from hybrid cells containing regions of the human genome in rodent cell backgrounds using primers directed to the human Alu repeat element. They demonstrate Alu-directed amplification of a fragment of the human HPRT gene from both hybrid cell and cloned DNA and identify through sequence analysis the Alu repeats involved in this amplification. They also demonstrate the application of this technique to identify the chromosomal locations of large fragments of the human X chromosome cloned in a yeast artificial chromosome and the general applicability of the method to the preparation of DNA probes from cloned human sequences. The technique allows rapid gene mapping and provides a simple method for the isolation and analysis of specific chromosomal regions.

• DOE Contract Number: FG05-88ER60692
• OSTI ID: 6968905
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 86:17; ISSN PNASA; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHROMOSOMAL ABERRATIONS
CHROMOSOMES
DAYS LIVING RADIOISOTOPES
DNA
DNA HYBRIDIZATION
DNA POLYMERASES
ELECTROPHORESIS
ENZYMES
GENE AMPLIFICATION
GENETIC MAPPING
HETEROCHROMOSOMES
HYBRIDIZATION
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAN
MAPPING
MOLECULAR BIOLOGY
MUTATIONS
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PRIMATES
RADIOISOTOPES
TRANSFERASES
VERTEBRATES
X CHROMOSOME