| pubmed-article:1616721 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:1616721 | lifeskim:mentions | umls-concept:C0020202 | lld:lifeskim |
| pubmed-article:1616721 | lifeskim:mentions | umls-concept:C0012854 | lld:lifeskim |
| pubmed-article:1616721 | lifeskim:mentions | umls-concept:C0002059 | lld:lifeskim |
| pubmed-article:1616721 | lifeskim:mentions | umls-concept:C1510438 | lld:lifeskim |
| pubmed-article:1616721 | lifeskim:mentions | umls-concept:C0303920 | lld:lifeskim |
| pubmed-article:1616721 | lifeskim:mentions | umls-concept:C1710236 | lld:lifeskim |
| pubmed-article:1616721 | pubmed:issue | 2 | lld:pubmed |
| pubmed-article:1616721 | pubmed:dateCreated | 1992-8-5 | lld:pubmed |
| pubmed-article:1616721 | pubmed:abstractText | A fluorometric procedure for the detection of DNA-DNA hybrids is described. The procedure involved the detection of probe-bound alkaline phosphatase with the fluorescent substrate ATTOPHOS. This substrate is converted to ATTOFLUOR by alkaline phosphatase and fluoresces strongly at 550 nm when excited with a wavelength of 440 nm. DNA hybridization assays were performed both with dilutions of purified target plasmid DNA (pSE9 or PBR322) and whole bacterial cells. Streptavidin-alkaline phosphatase conjugates were added to react with bound probe. Fluorometric assays, as well as colorimetric assays, using 5-bromo-4-chloro-3-indolylphosphate + nitroblue tetrazolium for alkaline phosphatase activity were performed. The fluorescence of the substrate was measured at time intervals, and the slope of the regression line calculated. A slope four times greater than that of background was considered positive. One hundred femtograms or 2.2 x 10(4) molecules of homologous DNA were detected with the fluorescent assay as compared with 10,000 femtograms or 2.2 x 10(6) molecules of homologous DNA with the colorimetric assay. Similar results were obtained with whole cells. Approximately 1 x 10(3) homologous cells were detected fluorometrically and 1 x 10(5) cells were detected colorimetrically. Based on these results, we conclude that, in our hands, the DNA hybridization assay described here using ATTOPHOS as the substrate for alkaline phosphatase is a very sensitive assay for the detection of DNA-DNA hybrids. | lld:pubmed |
| pubmed-article:1616721 | pubmed:language | eng | lld:pubmed |
| pubmed-article:1616721 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1616721 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:1616721 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1616721 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1616721 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1616721 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1616721 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1616721 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1616721 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:1616721 | pubmed:month | Feb | lld:pubmed |
| pubmed-article:1616721 | pubmed:issn | 0736-6205 | lld:pubmed |
| pubmed-article:1616721 | pubmed:author | pubmed-author:CaneR DRD | lld:pubmed |
| pubmed-article:1616721 | pubmed:author | pubmed-author:PalomaresJ... | lld:pubmed |
| pubmed-article:1616721 | pubmed:author | pubmed-author:TorresM JMJ | lld:pubmed |
| pubmed-article:1616721 | pubmed:author | pubmed-author:KlemR ERE | lld:pubmed |
| pubmed-article:1616721 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:1616721 | pubmed:volume | 12 | lld:pubmed |
| pubmed-article:1616721 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:1616721 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:1616721 | pubmed:pagination | 264-9 | lld:pubmed |
| pubmed-article:1616721 | pubmed:dateRevised | 2006-11-15 | lld:pubmed |
| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
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| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
| pubmed-article:1616721 | pubmed:meshHeading | pubmed-meshheading:1616721-... | lld:pubmed |
| pubmed-article:1616721 | pubmed:year | 1992 | lld:pubmed |
| pubmed-article:1616721 | pubmed:articleTitle | DNA hybridization assay using ATTOPHOS, a fluorescent substrate for alkaline phosphatase. | lld:pubmed |
| pubmed-article:1616721 | pubmed:affiliation | Biological Sciences Department, California Polytechnic University, San Luis Obispo 93407. | lld:pubmed |
| pubmed-article:1616721 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:1616721 | pubmed:publicationType | Comparative Study | lld:pubmed |
| pubmed-article:1616721 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:1616721 | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:1616721 | lld:pubmed |
