Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines

Journal Article · Sat Sep 01 04:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.87.17.6634· OSTI ID:5452229

Lichter, P; Ward, D C ^[1]; Ledbetter, S A; Ledbetter, D H ^[2]

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1. Yale Univ., New Haven, CT (USA)
2. Baylor College of Medicine, Houston, TX (USA)

Human-rodent hybrid cell lines have been analyzed with regard to their human DNA content by using various DNA probe sets, derived from the hybrids, for in situ hybridization to normal human metaphase chromosome spreads. Total genomic hybrid DNA was compared with probe sets of hybrid DNA that were highly enriched in human sequences. The latter probes were obtained by amplification through the polymerase chain reaction (PCR) using oligonucleotide primers directed to human specific subsequences of the interspersed repetitive sequences Alu and L1. Previously unidentified chromosomal material within hybrid lines was characterized with speed and precision. It is demonstrated that the complete human complement of hybrid lines can be rapidly assessed by comparing the data obtained with the Alu-PCR products with the results from the L1-PCR products or from the genomic hybrid DNA. This approach using interspersed repetitive sequence-PCR products is simple and fast and also provides an alternative way of generating complex DNA probe sets for the specific delineation of entire chromosomes or subchromosomal regions by the in situ hybridization.

• OSTI ID: 5452229
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 87:17; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
CHROMOSOMES
DNA HYBRIDIZATION
DNA POLYMERASES
DNA SEQUENCING
ENZYMES
FLUORESCENCE
GENE AMPLIFICATION
HYBRIDIZATION
LUMINESCENCE
MAMMALS
MAN
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PRIMATES
STRUCTURAL CHEMICAL ANALYSIS
TRANSFERASES
VERTEBRATES