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PMID: 2276735 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

"PCR-karyotype" of human chromosomes in somatic cell hybrids.

Genomics ·Vol. 8 ·No. 4 ·1990-12-00 ·Pages 614-22

Ledbetter SA, Garcia-Heras J, Ledbetter DH

Abstract

Amplification of human DNA sequences in 16 monochromosomal somatic cell hybrids containing different human chromosomes were performed by the polymerase chain reaction (PCR) using primer directed at human-specific regions of Alu or L1, the two major classes of interspersed repetitive sequences (IRS-PCR). A chromosome-specific pattern of amplification products was observed on agarose gels run with ethidium bromide, producing a "PCR-karyotype." This simple gel analysis provides a rapid method for identifying and monitoring the human chromosomal content of monochromosomal somatic cell hybrids without conventional cytogenetic analysis. Hybrids containing multiple human chromosome produce complex gel patterns, but identification of chromosome content can be achieved by hybridization of PCR products against a reference panel of monochromosomal or highly reduced hybrids representing each human chromosome. This dot-blot method also enables identification of human marker chromosomes or translocated pieces in hybrids that are not identifiable by cytogenetic methods. These IRS-PCR methods should greatly reduce the need for more laborious cytogenetic, isozyme, and Southern blot characterizations of human-rodent cell hybrids.

MeSH Terms
Animals Base Sequence Blotting, Southern Chromosomes, Human Cricetinae Electrophoresis, Agar Gel Humans Hybrid Cells Karyotyping/methods Molecular Sequence Data Polymerase Chain Reaction/methods Repetitive Sequences, Nucleic Acid Templates, Genetic
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ledbetter S A
Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
Garcia-Heras J
Ledbetter D H
Article Info
Journal
Genomics
Abbr.
Genomics
ISSN
0888-7543
Published
1990-12-00
Pages
614-22
Language
English
Region
United States
NLM ID
8800135
Subset
IM
Grants
NICHD NIH HHS · HD 20619 · United States
NICHD NIH HHS · HD 22478 · United States
NICHD NIH HHS · P30 HD24064 · United States
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