Detection of legumin gene DNA sequences in pea by in situ hybridization
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In order to detect low copy number sequences in pea using biotin-labelled probes we optimised some aspects of the in situ hybridization technique. We found protoplast preparations to be superior to standard squashes in terms of their signal: noise ratio. Heat and alkali denaturation of chromosomal DNA were both more effective than acid denaturation. A comparison of antibody-fluorochrome and streptavidin-enzyme conjugates showed the streptavidin-alkaline phosphatase conjugate to be the most sensitive detection system. Using the optimised method, we were able to detect a single site for a 13.5 kb legumin gene clone.
Originalsprog | Engelsk |
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Tidsskrift | Chromosoma |
Vol/bind | 96 |
Udgave nummer | 6 |
Sider (fra-til) | 454-458 |
Antal sider | 5 |
ISSN | 0009-5915 |
DOI | |
Status | Udgivet - jul. 1988 |
ID: 380059122