Optimized Detection of DNA Point Mutations by Double Gradient Denaturing Gradient Gel
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Laura Cremonesi
Abstract
Denaturing gradient gel electrophoresis displays the highest detection rate among mutation scanning methods. In classical denaturing gradient gel electrophoresis the denaturant gradient range and migration times vary for every amplicon to be scanned, greatly affecting the routine application of the method. As an alternative, we developed double gradient denaturing gradient gel electrophoresis where a gradient of pore size is superimposed over the denaturing one, allowing maintenance of the zone-sharpening effect even over prolonged time runs, and adoption of identical run time conditions for all fragments analyzed. Here double gradient denaturing gradient gel electrophoresis has been applied to the analysis of a number of point mutations and polymorphisms located in several exons of three different genes, the cystic fibrosis transmembrane conductance regulator, the β-globin and the p53 genes.
Copyright © 1999 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Author Index
- Contents
- Subject Index
- Polymorphisms of Coagulation Factor Genes a Review
- Urinalysis-Challenges by New Medical Needs and Advanced Technologies
- The Automation of Sediment Urinalysis Using a New Urine Flow Cytometer (UF-100™)
- Urinary Microscopy as Seen by Nephrologists
- Measurement of Urine Relative Density Using Refractometer and Reagent Strips
- Dry Chemistry Urinalysis of Pathological Proteinuria
- Physiopathology of Proteinuria and Laboratory Diagnostic Strategy Based on Single Protein Analysis
- Microalbuminuria in Diabetes
- European Multicentre Evaluation of the Super Aution SA-4220 Urinalysis Analyser
- Optimized Detection of DNA Point Mutations by Double Gradient Denaturing Gradient Gel
- Is the Association of Serum Lipase with β2-Microglobulin or C-Reactive Protein Useful for Establishing the Diagnosis and Prognosis of Patients with Acute Pancreatitis?
- Analytical and Clinical Performance of an Automated Immunoassay System (Immulite®) for Estradiol in Serum
- Evaluation of the Activated Partial Thrombo-plastin Time (APTT) Sensitivity to Heparin Using Five Commercial Reagents: Implications for Therapeutic Monitoring
- An Alternative Analysis for Crossover Studies that Accounts for Between-Group Disparities in Drug Response
Articles in the same Issue
- Author Index
- Contents
- Subject Index
- Polymorphisms of Coagulation Factor Genes a Review
- Urinalysis-Challenges by New Medical Needs and Advanced Technologies
- The Automation of Sediment Urinalysis Using a New Urine Flow Cytometer (UF-100™)
- Urinary Microscopy as Seen by Nephrologists
- Measurement of Urine Relative Density Using Refractometer and Reagent Strips
- Dry Chemistry Urinalysis of Pathological Proteinuria
- Physiopathology of Proteinuria and Laboratory Diagnostic Strategy Based on Single Protein Analysis
- Microalbuminuria in Diabetes
- European Multicentre Evaluation of the Super Aution SA-4220 Urinalysis Analyser
- Optimized Detection of DNA Point Mutations by Double Gradient Denaturing Gradient Gel
- Is the Association of Serum Lipase with β2-Microglobulin or C-Reactive Protein Useful for Establishing the Diagnosis and Prognosis of Patients with Acute Pancreatitis?
- Analytical and Clinical Performance of an Automated Immunoassay System (Immulite®) for Estradiol in Serum
- Evaluation of the Activated Partial Thrombo-plastin Time (APTT) Sensitivity to Heparin Using Five Commercial Reagents: Implications for Therapeutic Monitoring
- An Alternative Analysis for Crossover Studies that Accounts for Between-Group Disparities in Drug Response