Alu Polymerase Chain Reaction: A Method for Rapid Isolation of Human-Specific Sequences from Complex DNA Sources

• Published in: Proceedings of the National Academy of Sciences - PNAS 1989-09, Vol.86 (17), p.6686-6690
• Main Authors: Nelson, David L., Ledbetter, Susan A., Corbo, Laura, Victoria, Maureen F., Ramírez-Solis, Ramiro, Webster, Thomas D., Ledbetter, David H., Caskey, C. Thomas
• Format: Article
• Language: English
• Subjects: 550201 - Biochemistry- Tracer Techniques; ANIMALS; AUTORADIOGRAPHY; Bacteriophages; Base Sequence; BASIC BIOLOGICAL SCIENCES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; Biological and medical sciences; Cell lines; CHROMOSOMAL ABERRATIONS; Chromosome Mapping; CHROMOSOMES; Chromosomes, Human; Classical genetics, quantitative genetics, hybrids; Complementary DNA; DAYS LIVING RADIOISOTOPES; DNA; DNA - genetics; DNA - isolation & purification; DNA HYBRIDIZATION; DNA POLYMERASES; DNA-Directed DNA Polymerase; ELECTROPHORESIS; ENZYMES; Exons; Fundamental and applied biological sciences. Psychology; GENE AMPLIFICATION; Genes; Genetic Complementation Test; GENETIC MAPPING; Genetic vectors; Genetics of eukaryotes. Biological and molecular evolution; HETEROCHROMOSOMES; Human; Human chromosomes; Humans; Hybrid cells; Hybrid Cells - cytology; Hybridity; HYBRIDIZATION; Hypoxanthine Phosphoribosyltransferase - genetics; ISOTOPES; LIGHT NUCLEI; MAMMALS; MAN; MAPPING; MOLECULAR BIOLOGY; Molecular Sequence Data; MUTATIONS; NUCLEI; NUCLEIC ACIDS; NUCLEOTIDYLTRANSFERASES; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; PHOSPHORUS 32; PHOSPHORUS ISOTOPES; PHOSPHORUS-GROUP TRANSFERASES; Polymerase chain reaction; POLYMERASES; PRIMATES; RADIOISOTOPES; Thermus aquaticus; TRANSFERASES; VERTEBRATES; X CHROMOSOME
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.86.17.6686

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. We 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. We 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. We 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.