Single Strand Conformation

Polymorphisms (SSCP) Heteroduplex Mobility Assay (HMA)

Based on the following PRINCIPLES: Single strand molecules of DNA form 3D structures which are stabilized by weak intramolecular interactions.

The electrophoretic mobility that these molecules present in non-denaturing gels depends on the chain length and its conformations.

A single alteration in one base –like a mutation– can cause a conformational change of the DNA molecule which may be visualized by different electrophoretic mobility.

Based on the following PRINCIPLES: A heteroduplex is a double stranded molecule of nucleic acid created by the recombination of the compared single strands.

If there are differences –such as mutations– between the two single stranded chains, they form an irregular structure when recombined.

This non-regular fragments move slower in non-denaturing electrophoretic gels than regular ones.

METHODS FOR DETECTING MUTATIONS IN DNA PETNICA SCIENCE CENTER (Serbia, August 2012)

Anna Pujol Coma - Laia Gil Molas

OTHER TECHNIQUES NEEDED Isolation of DNA from bacteria, PCR, Agarose and polyacrylamide electrophoresis, Size-exclusion chromatography

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INTRODUCTION Since the discovering of the role of DNA in the development of life, scientists have been interested in the DNA mutations that can have an influence in our health. In our stay in Petnica, we worked in two techniques that allow researchers to detect mutations: Single Stranded Conformation Polymorphism (SSCP) and Heteroduplex mobility assay running. Our first goal was to optimize the SSCP technique for the new Petnica's laboratory, but the lack of time and some technical problems made us change our objective into the learning of the basic principles of some of the most well-known techniques in a molecular biology laboratory.

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