Alu-PCR: characterization of a chromosome 6-specific hybrid mapping panel and cloning of chromosome-specific markers.
The Alu-polymerase chain reaction (Alu-PCR) was applied to selectively amplify DNA sequences from human chromosome 6 using a single primer (A1) directed to the human Alu consensus sequence. A specific amplification pattern was demonstrated for a panel of eight somatic cell hybrids containing different portions of chromosome 6. This PCR pattern permits the identification of submicroscopic DNA alterations and can be utilized as a reference for additional chromosome 6-specific hybrids. To obtain new chromosome 6-specific markers we established two libraries from PCR-amplified sequences using two somatic cell hybrids (MCH381.2D and 640-5A). Out of a total of 109 clones that were found to be chromosome 6 specific, 13 clones were regionally assigned. We also included a procedure that allows the isolation of chromosome 6-specific markers from hybrids that contain human chromosomes other than 6. Our results will contribute to the molecular characterization of chromosome 6 by fostering characterization of somatic cell hybrids and by the generation of new regionally assigned DNA markers.
Meese
E U
EU
University of Michigan Medical Center, Department of Radiation Oncology, Ann Arbor 48109-0668.
Meltzer
P S
PS
Ferguson
P W
PW
Trent
J M
JM
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D002871:Chromosome Banding; D002896:Chromosomes, Human, Pair 6; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D016172:DNA Fingerprinting; D005819:Genetic Markers; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*
D005819:Genetic Markers; D004247:DNA