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231222s2000 xx ||||| 00| ||eng c |
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|a pubmed25n0364.xml
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|a (NLM)10997272
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|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Ekstrøm, P O
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Two-point fluorescence detection and automated fraction collection applied to constant denaturant capillary electrophoresis
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|c 2000
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|a Text
|b txt
|2 rdacontent
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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|a Band
|b nc
|2 rdacarrier
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|a Date Completed 18.01.2001
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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 Constant denaturant capillary electrophoresis (CDCE) has been shown to be a sensitive method to detect point mutations in DNA sequences of 100-bp lengths. Here, we report a significant modifications for the instrumental setup that allows a highly accurate prediction of the elution time of DNA fragments from the capillary and an efficient collection of separated fractions. Fluorescently labeled DNA fragments of TP53 exon 8 wild-type and two mutants (base pair number 14480 and 14525) are detected at two separate points of the same capillary. This permits the precise calculation of the fragment velocity after separation in the heated zone because, at room temperature, all DNA fragments of the same length have the same velocity. Such precision permits the selective collection of separated fragments using an automated fraction collector for additional CDCE analysis or sequencing. Also, the two-point detection allows one to rapidly distinguish between double-stranded and single-stranded DNA fragments of the same length, a process that cannot be achieved with a one-point detection system alone. Both modifications greatly improve the procedure to detect novel mutations by means of CDCE
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| 650 |
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|a Journal Article
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| 650 |
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|a Research Support, Non-U.S. Gov't
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| 650 |
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|a Research Support, U.S. Gov't, P.H.S.
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| 650 |
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7 |
|a Fluorescent Dyes
|2 NLM
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| 650 |
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7 |
|a Tumor Suppressor Protein p53
|2 NLM
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| 700 |
1 |
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|a Wasserkort, R
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Minarik, M
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Foret, F
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Thilly, W G
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t BioTechniques
|d 1993
|g 29(2000), 3 vom: 15. Sept., Seite 582-4, 586-9
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
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| 773 |
1 |
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|g volume:29
|g year:2000
|g number:3
|g day:15
|g month:09
|g pages:582-4, 586-9
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