ISSN:
1617-4623
Keywords:
Degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR)
;
Vicia faba Flow-sorted chromosomes
;
Southern hybridization Fluorescent in situ hybridization
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Degenerate oligonucleotide primed (DOP)-PCR has emerged as a simple and rapid method for representative amplification of highly complex genomic DNA from humans, mice and Drosophila. The present paper describes the adaptation of this method for use on a plant species, Vicia faba, with a large genome (2C = 30 pg). Specific low-copy-number sequences as well as highly repeated sequences were detectable among DOP-PCR products obtained from small samples of purified genomic DNA (100 pg), DNA from 10 prophase nuclei, 10 flow-sorted chromosomes or 15 microdissected chromosome segments (satellites) following reamplification with sequence-specific primers and/or Southern hybridization. Biotinylated chromosome-specific DOP-PCR products were used for fluorescent in situ hybridization. All chromosomes showed hybridization signals, with the exception of regions containing Fok elements which are not present in the chromosomal DNA targeted by DOP-PCR.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00280314
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