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PCR–SSCP: A Method for the Molecular Analysis of Genetic Diseases

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An Erratum to this article was published on 25 June 2008

Abstract

Single strand conformation polymorphism (SSCP) is a reproducible, rapid and quite simple method for the detection of deletions/insertions/rearrangements in polymerase chain reaction amplified DNA. All the details for the use of PCR–SSCP are presented in the direction of genetic diseases (β-thalassaemia, cystic fibrosis), optimum gel conditions, sensitivity and the latest modifications of the method, which are applied in most laboratories. This non-radioactive PCR–SSCP method can be reliably used to identify mutations in patients (β-globin, CFTR), provided suitable controls are available. Moreover, it is widely used for mutation identification in carriers (β-thalassaemia, cystic fibrosis), making it particularly useful in population screening.

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Abbreviations

ARMS:

Amplification refractory mutation system

ASO:

Allele specific oligonucleotide

Bi-ddF:

Bi-directional dideoxy fingerprinting

Bp:

Base pairs

CAE-SSCP:

Capillary array electrophoresis–SSCP

CE-SSCPC:

Capillary electrophoresis–SSCP

CF:

Cystic fibrosis

CFTR :

Cystic fibrosis transmembrane regulator

DGGE:

Denaturing gradient gel electrophoresis

DHPLC:

Denaturing high-performance liquid chromatography

DNTP:

Deoxynucleoside triphosphates

EDTA:

Ethylene diamine tetra-acetic acid

HD:

Heteroduplex

IVS:

Intervening sequence

PAGE:

Polyacrilymide gel electrophoresis

PCR:

Polymerase chain reaction

RFLP:

Restriction fragment length polymorphism

SDS:

Sodium dodecyl sulphate

SSCP:

Single strand conformational polymorphism

ssDNA or dsDNA:

Single stranded or double stranded DNA

TPE:

Tris phosphate EDTA

TBE:

Tris borate EDTA

TGGE:

Temperature gradient gel electrophoresis

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Acknowledgements

We thank Associate Professor S. Bonanou, for many helpful suggestions and help in 2003 and Ms T. Tsianou for her valuable contribution to the CFTR project. Grand sponsor European Union—Archimedes II.

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  1. Laboratory of Clinical Chemistry, School of Medical Laboratories, Faculty of Health and Care, Highest Technological Institute of Larissa, 37 Xenopulu str, Larissa, 41221, Greece

    Kakavas V. Konstantinos, Plageras Panagiotis, Vlachos T. Antonios, Papaioannou Agelos & Noulas V. Argiris

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  1. Kakavas V. Konstantinos
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Correspondence to Kakavas V. Konstantinos.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s12033-008-9077-0

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Konstantinos, K.V., Panagiotis, P., Antonios, V.T. et al. PCR–SSCP: A Method for the Molecular Analysis of Genetic Diseases. Mol Biotechnol 38, 155–163 (2008). https://doi.org/10.1007/s12033-007-9006-7

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