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

Masabanda, Julio S, Griffin, Darren K. (2003) Generation of chromosome paints: approach for increasing specificity and intensity of signals. BioTechniques, 34 (3). 530-2, 534, 536. ISSN 0736-6205. (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:61070)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided.

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
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Susan Davies
Date Deposited: 28 Mar 2017 14:15 UTC
Last Modified: 16 Nov 2021 10:24 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/61070 (The current URI for this page, for reference purposes)

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