1. METHODS FOR DETECTING MUTATIONS IN DNA
PETNICA SCIENCE CENTER (Serbia, August 2012)
Anna Pujol Coma - Laia Gil Molas
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.
Single Strand Conformation
Heteroduplex Mobility Assay (HMA)
Polymorphisms (SSCP)
Based on the following PRINCIPLES: Based on the following PRINCIPLES:
 Single strand molecules of DNA form 3D structures which  A heteroduplex is a double stranded molecule of nucleic
are stabilized by weak intramolecular interactions. acid created by the recombination of the compared single
 The electrophoretic mobility that these molecules present strands.
in non-denaturing gels depends on the chain length and  If there are differences –such as mutations– between the
its conformations. two single stranded chains, they form an irregular
 A single alteration in one base –like a mutation– can cause structure when recombined.
a conformational change of the DNA molecule which may  This non-regular fragments move slower in non-
be visualized by different electrophoretic mobility. denaturing electrophoretic gels than regular ones.
OTHER TECHNIQUES NEEDED
Isolation of DNA from bacteria, PCR, Agarose and polyacrylamide electrophoresis, Size-exclusion chromatography
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