A mouse Chromosome 11 library generated from sorted chromosomes using linker-adapter polymerase chain reaction

K. Miyashita, M. A. Vooijs, J. D. Tucker, D. A. Lee, J. W. Gray, Maria G. Pallavicini

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

We describe the generation of a mouse whole chromosome library using sequence-independent polymerase chain reaction (PCR) to amplify sequences contained in DNA extracted from flow sorted chromosomes. DNA in sorted chromosomes from a human × mouse hybrid cell line was digested with a frequent four-cutter restriction enzyme. Sau3AI, and the ends were ligated to an adapter oligonucleotide. The ligated DNA fragments were amplified using PCR primers homologous to the linker-adapter oligonucleotide. PCR-generated products were characterized by gel electrophoresis. The size of the amplified DNA ranged from 100 to more than 1000 bp with a relatively high proportion of products at approximately 400 bp. Biotinylated PCR products used for FISH showed specific hybridization to murine metaphases and no hybridization to human lymphocyte and hamster metaphase cells. Banding analysis indicated that the probes were specific for mouse Chromosome 11. We anticipate that availability of painting probes for specific murine chromosomes will facilitate cytogenetic studies in the mouse.

Original languageEnglish (US)
Pages (from-to)54-57
Number of pages4
JournalCytogenetic and Genome Research
Volume66
Issue number1
DOIs
StatePublished - Jan 1 1994

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

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