Multiplexed p53 Mutation Detection by Microchip Electro-phoresis with Laser-Induced Fluorescence Detector
A form of single‐strand DNA‐conformation polymorphism analysis (SSCP) employing nondenaturing slab gel electrophoresis is applicable to the genetic diagnosis of mutations at exons 7, 8 and 9 of the p53 gene. Recently, microchip electrophoresis (ME) systems have been used in SSCP analysis instead of...
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| Published in: | Chinese journal of chemistry Vol. 28; no. 5; pp. 797 - 802 |
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| Main Authors: | , , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
Weinheim
WILEY-VCH Verlag
01-05-2010
WILEY‐VCH Verlag Wiley Subscription Services, Inc |
| Subjects: | |
| Online Access: | Get full text |
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| Summary: | A form of single‐strand DNA‐conformation polymorphism analysis (SSCP) employing nondenaturing slab gel electrophoresis is applicable to the genetic diagnosis of mutations at exons 7, 8 and 9 of the p53 gene. Recently, microchip electrophoresis (ME) systems have been used in SSCP analysis instead of conventional slab gel electrophoresis in terms of speed, sensitivity and automation. The aim of the present study was to investigate the application of SSCP and ME analysis as a rapid and effective method to the detection of mutations for exons 7, 8 and 9 of the p53 gene. It was found that using the electric field strength 260 V/cm and the sieving matrix of 4 mg/mL poly(ethylene oxide) was very useful to achieve better resolution and fast detection of mutations at exons 7, 8 and 9 of p53 gene. Under the optimized conditions, mutations at exons 7–9 of p53 gene were analyzed within 60 s and the relative standard deviation values of the migration times were less than 5.81% (n=5). The detection limit can be as low as 1 ng·L−1.
Schematic diagram illustrating the general principles of the ME‐SSCP: Reservoir 1=buffer inlet, reservoir 2=sample outlet, reservoir 3=buffer waste, reservoir 4=sample inlet. |
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| Bibliography: | Science and Technology Commission of Shanghai - No. 08142201400 ark:/67375/WNG-N42N8JX2-L istex:7BCEA7B875F41177C913A64A60103289772053F8 ArticleID:CJOC201090149 New Century Excellent Talents in University ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
| ISSN: | 1001-604X 1614-7065 |
| DOI: | 10.1002/cjoc.201090149 |