Alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex DNA sources.
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.
Nelson
D L
DL
Howard Hughes Medical Institute, Baylor College of Medicine, Houston 77030.
Ledbetter
S A
SA
Corbo
L
L
Victoria
M F
MF
Ramírez-Solis
R
R
Webster
T D
TD
Ledbetter
D H
DH
Caskey
C T
CT
D001483:Base Sequence; D002874:Chromosome Mapping; D002877:Chromosomes, Human; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D005796:Genes; D005816:Genetic Complementation Test; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008969:Molecular Sequence Data
D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase; D004259:DNA-Directed DNA Polymerase