GrainGenes Reference Report: GNM-36-701
Reference
GNM-36-701
Title
An optimized fluorescence in situ hybridization procedure for detecting rye chromosomes in wheat
Journal
Genome
Year
1993
Volume
36
Pages
701-705
Author
Nkongolo K Lapitan N Quick J Muhlmann M
Abstract
In situ hybridization with an interspersed repeat clone from rye, pSc119, was shown to be useful for detecting rye chromosomes introduced into wheat However, since pSc119 also shows strong hybridization to a few sites in certain wheat chromosomes, small rye chromosome segments added to wheat may be difficult to detect In this study, detection of rye chromosomes present in triticale and triticale x wheat hybrids was accomplished with the use of a subfragment from pSc119 (pSc119.1) whose sequence is dispersed throughout the rye chromosomes and only weakly cross-hybridizes to a few telomeric and centromeric regions of wheat. The in situ hybridization conditions were optimized to readily distinguish rye chromosomes from wheat chromosomes without the need for intensive analysis of hybridization patterns. Rye chromosomes were readily detected using fluorescence in situ hybridization. Fluorescence detection provided increased sensitivity over enzymatic detection and allowed signals to be amplified with repeated use of biotinylated anti-avidin antibody and avidin-exposure FITC. Detection of rye chromatin was further optimized by doubling the probe concentration. Finally, double exposure photography of the same cell with two different filters provided another means to further increase the contrast between rye and wheat chromosomes.
Keyword
chromosome
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GrainGenes Reference Report: GNM-36-701
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GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture. | |||