How to cite this article: Lucas JN, Wu X, Guo E, Chi LE, Chen Z. 2006. An efficient chemical method to generate repetitive sequences depleted DNA probes. Am J Med Genet Part A 140A:2115–2120.
1552-4833/asset/olbannerleft.gif?v=1&s=dc5513758e0602016bfe88cd52afd78bf862aab9)
1552-4833/asset/olbannerright.gif?v=1&s=c3d8a0f0a45667625907578ec1e0f011cbe060b1)
Research Article
You have free access to this content
An efficient chemical method to generate repetitive sequences depleted DNA probes†
Article first published online: 12 JUL 2006
DOI: 10.1002/ajmg.a.31327
Copyright © 2006 Wiley-Liss, Inc.
Issue
1552-4833/asset/cover.gif?v=1&s=9818ec8c89d0772bf0e752b15694c460afb83f9f "American Journal of Medical Genetics Part A")
American Journal of Medical Genetics Part A
Special Issue: John M. Opitz Festschrift
Volume 140A, Issue 19, pages 2115–2120, 1 October 2006
Additional Information
How to Cite
Lucas, J. N., Wu, X., Guo, E., Chi, L. E. and Chen, Z. (2006), An efficient chemical method to generate repetitive sequences depleted DNA probes. American Journal of Medical Genetics Part A, 140A: 2115–2120. doi: 10.1002/ajmg.a.31327
- †
Publication History
- Issue published online: 22 SEP 2006
- Article first published online: 12 JUL 2006
- Manuscript Accepted: 25 APR 2006
- Manuscript Received: 1 DEC 2005
Funded by
- NIH. Grant Number: R43 CA117161-01
- DOD. Grant Number: DAMD17-02-C-0008
- Abstract
- Article
- References
- Cited By
Keywords:
- repetitive sequences;
- chemical method;
- human DNA;
- FISH;
- subtraction;
- phenol
Abstract
We describe an efficient method to remove repetitive sequences from DNA of microdissected whole chromosomes, chromosome arms, locus specific probes, and specific bands. The chemical approach described removes human repetitive DNA sequences from a source DNA, eliminating the need for blocking such sequences when the source DNA used as a probe is hybridized with a target DNA. It employs subtracting hybridized biotin-labeled repetitive-sequence DNA complex with phenol and chloroform after incubation of hybridized products with avidin. The method produces unique products that are formed after such repetitive sequences have been removed from the DNA. We have applied our newly designed subtraction strategy to microdissected chromosomes in generating whole chromosome painting (e.g., chromosome 4), chromosome arm (e.g., 1q and 15q), and band (e.g., 3q26) specific probes, and we have demonstrated its utility using flow-sorted chromosome. FISH analyses using these probes show consistent strong signals with no cross-hybridization, and optimal hybridization results can be obtained relatively quickly. © 2006 Wiley-Liss, Inc.