Fingerprinting human chromosomes by polymerase chain reaction-mediated DNA amplification.
We describe here a method for DNA fingerprinting of human chromosomes by Alu-polymerase chain reaction (PCR) amplification of DNA from monochromosomal hybrids, following digestion with restriction endonucleases. DNA digestion with restriction enzymes prior to PCR amplification reduces the total number of amplified fragments. The number and pattern of bands of PCR products observed in an electrophoretic medium are chromosome specific and provide a "fingerprint signature" for individual human chromosomes. Using this approach, we have produced fingerprints for human chromosomes 2, 5, 7, 9, and 12. The applicability of this approach to chromosome identification was assessed by comparing the fingerprints obtained for two different hybrids containing chromosome 7. DNA fragments specific for the long and the short arms of human chromosome 12 have also been identified. In addition, Alu-PCR-generated DNA fragments, specific for different chromosomes, were used to probe Southern blots of a hybrid cell panel to identify human chromosomes present in hybrid cell lines. The chromosomal specificity of these probes permits the identification of intact as well as rearranged chromosomes composed of segments arising from more than one chromosome.
Sidhu
M S
MS
Department of Microbiology and Molecular Genetics, New Jersey Medical School, Newark 07103.
Helen
B K
BK
Athwal
R S
RS
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002877:Chromosomes, Human; D006224:Cricetinae; D016172:DNA Fingerprinting / Q000379:methods*; D004277:DNA, Single-Stranded; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid
D004277:DNA, Single-Stranded