An apparatus for submerged gel electrophoresis

Kozulić, Branko; Heimgartner, Urs

doi:10.1016/0003-2697(91)90422-p

Cited by 6 publications

(2 citation statements)

References 5 publications

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“…Spreadex gels are made by free radical polymerization. The gel polymers are arranged in a unique 3-D structure, in which they provide increased resistance to the migrating DNA molecules in accordance with theoretical considerations on gel electrophoresis (1,7,8). The Spreadex gels are characterized by an exclusion limit such that DNA fragments with sizes above the specific limit are prevented from migrating into the gel.…”

Section: Introductionmentioning

confidence: 78%

High-Resolution Detection of Loss of Heterozygosity of Dinucleotide Microsatellite Markers

2001

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Dinucleotide microsatellite markers are frequently investigated to study inheritance, genetic stability, and allele frequency distribution in a wide variety of genetic disorders. Previous studies have encountered significant problems regarding resolution and detection of dinucleotide, microsatellites. In this study, a useful method to investigate loss of heterozygosity (LOH) of dinucleotide microsatellite markers is described that involves the use of nondenaturing (Spreadex) submerged gel electrophoresis and SYBR Green I nucleic acid staining. This method omits the gel casting step and the use of hazardous radioactive materials frequently used in many microsatellite studies that employ polyacrylamide gel nucleic acid denaturation analysis. Using this method, 62 patients' paired tumor and normal samples were investigated to detect allele deletions in a region of chromosome 7q31.1, which is believed to harbor a tumor suppressor gene. Interpretable results were obtained in all cases. These results were compared to those attained using ABI Prism Genetic Analyzer 310 and Gene-Scan. There were no discrepancies in results obtained between the two assays. The Spreadex system is cheap, does not require larger equipment costs, and may prove to be a useful system for high-throughput investigation of microsatellites. It may have diagnostic significance and also prove useful if applied to population-based genomic screening and linkage analysis.

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Section: Introductionmentioning

confidence: 78%

High-Resolution Detection of Loss of Heterozygosity of Dinucleotide Microsatellite Markers

2001

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“…On the other hand, the SEA 2000 submarine electrophoresis system is provided with a heat exchange element and a pump for laminar buffer flow. These elements are specially designed to maintain the uniform temperature level all over the gel surface (24). Because the SSCP pattern reproducibility depends largely on temperature, temperature control is essential.…”

Section: Discussionmentioning

confidence: 99%

Detection of Factor V Leiden by PCR-SSCP using GMATM Precast Elchrom Scientific Gels

Šimundić¹,

Topić²,

Štefanović³

2003

Clin Appl Thromb Hemost

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Genetic abnormalities in hemostatic proteins associated with hypercoagulability are an important hereditary risk factor for venous thrombosis. Several genetic mutations that cause hereditary disorders predisposing to thrombosis have been described, point mutation in the coagulation factor V gene (FV:R506Q), called factor V Leiden, being the most common of them. A new inexpensive and simple polymerase chain reaction-single-strand polymorphism (PCR-SSCP) based method for detection of this genetic abnormality is reported. The study population consisted of 150 subjects whose factor V genotype was previously determined by PCR-RFLP method using the Mnl I restriction endonuclease. A 223-bp fragment containing the G1692-A (Arg 506-Gln) polymorphic site in exon 10 of the factor V gene was amplified, denatured, and run overnight on the commercially available GMA gels for SSCP. PCR-SSCP analysis showed reproducible and uniform band patterns for FV mutant and wild type alleles. Furthermore, PCR-SSCP results were consistent with those obtained with PCR-RFLP analysis (100%). The described PCR-SSCP procedure is reliable, time-saving, and cost-effective. The method may be considered as a potentially powerful new tool in the routine detection of factor V Leiden.

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