Catch-linker + PCR labeling: a simple method to generate fluorescence in situ hybridization probes from yeast artificial chromosomes. | Zendy

Yoshiro Shibasaki | Zendy; John C. Maule | Zendy; Rebecca S. Devon | Zendy; Euan M. Slorach | Zendy; John R. Gosden | Zendy; David J. Porteous | Zendy; Anthony J. Brookes | Zendy

Open Access

Catch-linker + PCR labeling: a simple method to generate fluorescence in situ hybridization probes from yeast artificial chromosomes.

Author(s) -

Yoshiro Shibasaki,

John C. Maule,

Rebecca S. Devon,

Euan M. Slorach,

John R. Gosden,

David J. Porteous,

Anthony J. Brookes

Publication year - 1995

Publication title -

genome research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.556

H-Index - 297

eISSN - 1549-5469

pISSN - 1088-9051

DOI - 10.1101/gr.4.4.209

Subject(s) - biology , fluorescence in situ hybridization , yeast , in situ , genetics , linker , microbiology and biotechnology , in situ hybridization , yeast artificial chromosome , fluorescence , computational biology , chromosome , gene mapping , gene , messenger rna , optics , physics , meteorology , computer science , operating system

A simple and efficient method to generate hapten-labeled DNA fragments from a trace amount of YAC DNA isolated by PFGE is described. After agarase digestion of the gel slice containing the resolved YAC recombinant, the purified DNA is digested with Sau3Al and a compatible CL oligonucleotide duplex ligated on. A probe is generated by PCR amplification using a primer complementary to the CL with a single biotin moiety incorporated at the 5' end. When used as a FISH probe, this material yields mapping results superior to Alu-PCR or whole YAC labeling methods and allows sensitive detection of chimerism.

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