Generation of chromosome paints: approach for increasing specificity and intensity of signals

Masabanda, Julio S. and Griffin, Darren K. (2003) Generation of chromosome paints: approach for increasing specificity and intensity of signals. BioTechniques, 34 (3). pp. 530-545. ISSN 0736-6205 . (The full text of this publication is not available from this repository)

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Abstract

Chromosome painting is a widely used technique, and the two principal means of generating probes for such experiments involve DNA isolation by chromosome flow sorting and by chromosome microdissection. Frequently, chromosome paints are bright and specific; however, on occasion, signals can be weak and nonspecific, particularly for microdissected probes. Reasons for this have been attributed to co-amplification of non-target DNA and the formation of primer concatamers during degenerate oligonucleotide primed (DOP)-PCR. Here we describe a technique of circumventing this problem by sequence enrichment. It involves co-hybridization of DOP-PCR biotinylated microdissected material and linkered genomic DNA. Biotinylated DNA fragments captured on streptavidin-coated paramagnetic beads are eluted and amplified by PCR using a single primer complementary to the linker arm.

Item Type: Article
Additional information: 12661159 0736-6205
Uncontrolled keywords: Animals Chickens Chromosome Painting Chromosomes, Mammalian Cricetinae In Situ Hybridization, Fluorescence Microscopy, Fluorescence Polymerase Chain Reaction Quality Control Reproducibility of Results Sensitivity and Specificity
Subjects: Q Science
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: Darren Griffin
Date Deposited: 10 Sep 2008 12:46
Last Modified: 21 May 2014 12:58
Resource URI: http://kar.kent.ac.uk/id/eprint/12480 (The current URI for this page, for reference purposes)
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