Abstract
A simple and fast method with high reliability is necessary for the identification of mutations, polymorphisms and sequence variants (MPSV) within many genes and many samples, e.g. for clarifying the genetic background of individuals with multifactorial diseases. Here we review our experience with the polymerase chain reaction/single-strand conformation polymorphism (PCR/SSCP) analysis to identify MPSV in a number of genes thought to be involved in the pathogenesis of multifactorial neurological disorders, including autoimmune diseases like multiple sclerosis (MS) and neurodegenerative disorders like Parkinsons disease (PD). The method is based on the property of the DNA that the electrophoretic mobility of single stranded nucleic acids depends not only on their size but also on their sequence. The target sequences were amplified, digested into fragments ranging from 50-240 base pairs (bp), heat-denatured and analysed on native polyacrylamide (PAA) gels of different composition. The analysis of a gre at number of different PCR products demonstrates that the detection rate of MPSV depends on the fragment lengths, the temperature during electrophoresis and the composition of the gel. In general, the detection of MPSV is neither influenced by their location within the DNA fragment nor by the type of substitution, i.e., transitions or transversions. The standard PCR/SSCP system described here provides high reliability and detection rates. It allows the efficient analysis of a large number of DNA samples and many different genes.
Keywords: PCR/SSCP, DNA sequence ariations, Base pairs, Parkinsons disease, Polyacrylamide
Combinatorial Chemistry & High Throughput Screening
Title: PCR/SSCP Detects Reliably and Efficiently DNA Sequence Variations in Large Scale Screening Projects
Volume: 3 Issue: 3
Author(s): Bianca Miterski, Rejko Kruger, Philip Wintermeyer and Jorg T. Epplen
Affiliation:
Keywords: PCR/SSCP, DNA sequence ariations, Base pairs, Parkinsons disease, Polyacrylamide
Abstract: A simple and fast method with high reliability is necessary for the identification of mutations, polymorphisms and sequence variants (MPSV) within many genes and many samples, e.g. for clarifying the genetic background of individuals with multifactorial diseases. Here we review our experience with the polymerase chain reaction/single-strand conformation polymorphism (PCR/SSCP) analysis to identify MPSV in a number of genes thought to be involved in the pathogenesis of multifactorial neurological disorders, including autoimmune diseases like multiple sclerosis (MS) and neurodegenerative disorders like Parkinsons disease (PD). The method is based on the property of the DNA that the electrophoretic mobility of single stranded nucleic acids depends not only on their size but also on their sequence. The target sequences were amplified, digested into fragments ranging from 50-240 base pairs (bp), heat-denatured and analysed on native polyacrylamide (PAA) gels of different composition. The analysis of a gre at number of different PCR products demonstrates that the detection rate of MPSV depends on the fragment lengths, the temperature during electrophoresis and the composition of the gel. In general, the detection of MPSV is neither influenced by their location within the DNA fragment nor by the type of substitution, i.e., transitions or transversions. The standard PCR/SSCP system described here provides high reliability and detection rates. It allows the efficient analysis of a large number of DNA samples and many different genes.
Export Options
About this article
Cite this article as:
Miterski Bianca, Kruger Rejko, Wintermeyer Philip and Epplen T. Jorg, PCR/SSCP Detects Reliably and Efficiently DNA Sequence Variations in Large Scale Screening Projects, Combinatorial Chemistry & High Throughput Screening 2000; 3 (3) . https://dx.doi.org/10.2174/1386207003331607
DOI https://dx.doi.org/10.2174/1386207003331607 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Synthesis, Antioxidant, Anticancer and Antiviral Activities of Novel Quinoxaline Hydrazone Derivatives and their Acyclic C-Nucleosides
Medicinal Chemistry Direct Targeting of the Ras GTPase Superfamily Through Structure- Based Design
Current Topics in Medicinal Chemistry Gabapentin-Mediated Effects on Voltage- and Ligand-Gated Currents
Current Neuropharmacology Recent Progresses in Application of Fullerenes in Cosmetics
Recent Patents on Biotechnology Pharmacology and Toxicology of Cannabis Derivatives and Endocannabinoid Agonists
Recent Patents on CNS Drug Discovery (Discontinued) Targeting Heme Oxygenase-1 for Neuroprotection and Neuroinflammation in Neurodegenerative Diseases
Current Drug Targets Beneficial Effects of Dietary Supplements Against Mitochondrial Dysfunction and Apoptosis in Neurodegenerative Diseases
Current Pharmacogenomics and Personalized Medicine Dysregulation of SIRT-1 Signaling in Multiple Sclerosis and Neuroimmune Disorders: A Systematic Review of SIRTUIN Activators as Potential Immunomodulators and their Influences on other Dysfunctions
Endocrine, Metabolic & Immune Disorders - Drug Targets Flavonoids in the Treatment of Diabetes: Clinical Outcomes and Mechanism to Ameliorate Blood Glucose Levels
Current Diabetes Reviews Bridge Between Neuroimmunity and Traumatic Brain Injury
Current Pharmaceutical Design The Molecular Basis of Conantokin Antagonism of NMDA Receptor Function
Current Drug Targets Recent Patents on Light Based Therapies: Photodynamic Therapy, Photothermal Therapy and Photoimmunotherapy
Recent Patents on Endocrine, Metabolic & Immune Drug Discovery Anti-Inflammatory Activity of Tetracyclines: Applications to Human Disease
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Fluorinated Molecules as Drugs and Imaging Agents in the CNS
Current Topics in Medicinal Chemistry Review of Theoretical Studies for Prediction of Neurodegenerative Inhibitors
Mini-Reviews in Medicinal Chemistry Alzheimer’s Disease-like Early-phase Brain Pathogenesis: Self-curing Amelioration of Neurodegeneration from Pro-inflammatory ‘Wounding’ to Anti-inflammatory ‘Healing’
Current Alzheimer Research Possible Neuroprotective Strategies in ALS
Current Neuropharmacology From the Editors Perspective: Inflammatory Glial Cells of the Nervous System: Assistants or Assassins?
Current Neurovascular Research The Protective Effects of Crocetin on Aβ<sub>1-42</sub>-Induced Toxicity in Ht22 Cells
CNS & Neurological Disorders - Drug Targets Antioxidants Suppress Th1-Type Immune Response In Vitro
Drug Metabolism Letters