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      Sensitivity and applications of the PCR Single-Strand Conformation Polymorphism method

      review-article
      Molecular Biology Reports
      Springer Netherlands
      SSCP, Applications, Coronavirus

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          Abstract

          PCR Single-Strand Conformation Polymorphism is a method used to identify and detect mutations and is now well known for its many applications on living beings. This paper will discuss the experimental details, limitations and sensitivity of the PCR Single-Strand Conformation Polymorphism method in relation to all existing literature available to us until today. Genomic DNA extraction, PCR amplification and Single-Strand Conformation Polymorphism conditions (concentration of polyacrylamide slab gel electrophoresis, dissociation treatment of double- stranded DNA) and comparison with PCR Restriction Fragment Length Polymorphism are presented. Since its discovery in 1989, there have been many variations, innovations, and modifications of the method, which makes it very easy, safe, fast and for this reason widely applied in clinical diagnostic, forensic medicine, biochemical, veterinary, microbiological, food and environmental laboratories. One of the possible applications of the method is the diagnosis and identification of mutations in new strains of coronaviruses, because science needs more tools to tackle the problem of this pandemic. The PCR Single-Strand Conformation Polymorphism method can be applied in many cases provided that control samples are available and the required conditions of the method are achieved.

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          Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction

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            SSCP is not so difficult: the application and utility of single-stranded conformation polymorphism in evolutionary biology and molecular ecology.

            All genetic markers are estimators of DNA nucleotide sequence variation. Rather than obtaining DNA sequence data, it is cheaper and faster to use techniques that estimate sequence variation, although this usually results in the loss of some information. SSCP (single-stranded conformation polymorphism) offers a sensitive but inexpensive, rapid, and convenient method for determining which DNA samples in a set differ in sequence, so that only an informative subset need be sequenced. In short, most DNA sequence variation can be detected with relatively little sequencing. SSCP has been widely applied in medical diagnosis, yet few studies have been published in population genetics. The utility and convenience of SSCP is far from fully appreciated by molecular population biologists. We hope to help redress this by illustrating the application of a single simple SSCP protocol to mitochondrial genes, nuclear introns, microsatellites, and anonymous nuclear sequences, in a range of vertebrates and invertebrates.
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              How sensitive is PCR-SSCP?

              Single-strand conformation polymorphism analysis (SSCP) is a rapid method for detection of minor sequence changes in polymerase chain reaction-amplified DNA. Since the first reported use of SSCP in 1989 (Orita et al., 1989), this technique has been used widely to detect mutations in oncogenes, tumor suppressor genes, and genes responsible for genetic diseases. Published mutations that have been detected using this technique include base substitutions, small insertions and deletions, and rearrangements. This technique has also been applied for the detection of DNA polymorphisms at various loci of the human genome (reviewed by Hayashi, 1991; Hayashi, 1993). However, many factors can influence the sensitivity of SSCP, and its optimization is highly empirical. In this review, we estimate the percentage of mutations that can be detected by this technique under various controlled conditions, and describe some critical elements affecting sensitivity.
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                Author and article information

                Contributors
                kakavas@uth.gr
                Journal
                Mol Biol Rep
                Mol Biol Rep
                Molecular Biology Reports
                Springer Netherlands (Dordrecht )
                0301-4851
                1573-4978
                24 April 2021
                : 1-7
                Affiliations
                GRID grid.410558.d, ISNI 0000 0001 0035 6670, Laboratory of Analytical, Environmental Chemistry and Human Health, Department of Environmental sciences, , University of Thessaly, Geopolis, ; 41500 Larisa, Greece
                Author information
                http://orcid.org/0000-0001-8247-0134
                Article
                6349
                10.1007/s11033-021-06349-2
                8065318
                33893925
                ac83604a-ed5e-4a87-a677-3cfc75f15825
                © The Author(s), under exclusive licence to Springer Nature B.V. 2021

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                : 5 January 2021
                : 8 April 2021
                Categories
                Review

                Molecular biology
                sscp,applications,coronavirus
                Molecular biology
                sscp, applications, coronavirus

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