Libraries for each human chromosome, constructed from sorter-enriched chromosomes by using linker-adaptor PCR

M. Vooijs, L. C. Yu, D. Tkachuk, D. Pinkel, D. Johnson, Joe Gray

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

We describe here the production of complex libraries enriched in sequences from each human chromosome type, starting with only a few thousand sorter-purified chromosomes. In this procedure, DNA is extracted from the sorted chromosomes, digested to completion by using the frequently cutting restriction endonuclease Sau3A1, and ligated, on each end, to an adaptor oligonucleotide. These fragments are then amplified using PCR with a sequence homologous to the adaptor oligonucleotide as a primer. We have used this procedure to produce PCR libraries for each of the 24 human chromosomes. These libraries were characterized by gel electrophoresis and found to be composed of a continuum of sequences ranging in size from a few hundred to ∼ 1,000 bp. The libraries, when used as probes for fluorescence in situ hybridization, stained the target chromosomes more or less continuously, even after PCR amplification for more than 200 cycles. These libraries are useful as hybridization probes to facilitate molecular cytogenetic studies and as sources of probes either for identification of polymorphic short tandemly repeated sequences or for development of sequence-tagged sites.

Original languageEnglish (US)
Pages (from-to)586-597
Number of pages12
JournalAmerican Journal of Human Genetics
Volume52
Issue number3
StatePublished - Mar 1993
Externally publishedYes

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Human Chromosomes
Libraries
Chromosomes
Polymerase Chain Reaction
Oligonucleotides
Sequence Tagged Sites
DNA Restriction Enzymes
Sequence Homology
Fluorescence In Situ Hybridization
Cytogenetics
Electrophoresis
Gels
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

Libraries for each human chromosome, constructed from sorter-enriched chromosomes by using linker-adaptor PCR. / Vooijs, M.; Yu, L. C.; Tkachuk, D.; Pinkel, D.; Johnson, D.; Gray, Joe.

In: American Journal of Human Genetics, Vol. 52, No. 3, 03.1993, p. 586-597.

Research output: Contribution to journalArticle

Vooijs, M. ; Yu, L. C. ; Tkachuk, D. ; Pinkel, D. ; Johnson, D. ; Gray, Joe. / Libraries for each human chromosome, constructed from sorter-enriched chromosomes by using linker-adaptor PCR. In: American Journal of Human Genetics. 1993 ; Vol. 52, No. 3. pp. 586-597.
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