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|a eng
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|a Cremonesi, L
|e verfasserin
|4 aut
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|a Double-gradient DGGE for optimized detection of DNA point mutations
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|c 1997
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|a Text
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|a ohne Hilfsmittel zu benutzen
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|a Date Completed 02.05.1997
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|a Date Revised 28.09.2018
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|a published: Print
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|a Citation Status MEDLINE
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|a A novel technique is reported for screening point mutations is genomic DNA: double gradient, denaturing gradient gel electrophoresis (DG-DGGE). Unlike conventional DGGE, which exploits a single gradient of denaturing chemicals (typically urea and formamide) along the migration path to force the two hetero- and two homo-duplexes to partially unwind and separate, DG-DGGE superimposes a second (porous) gradient over the denaturing one. With the help of the sieving gradient, molecules such as the hetero-duplexes, which often produce curtains and smears instead of sharp zones, due to lack of a sharp melting transition, are re-compacted into remarkably narrow bands. Even homo-duplexes with minute melting temperature differences, giving a single band in DGGE, are resolved into two zones in DG-DGGE. The technique has been applied to the analysis of a number of point mutations in several exons of the cystic fibrosis transmembrane conductance regulator gene
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|a Gelfi, C
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|i Enthalten in
|t BioTechniques
|d 1991
|g 22(1997), 2 vom: 31. Feb., Seite 326-30
|w (DE-627)NLM012627046
|x 1940-9818
|7 nnas
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| 773 |
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