Optimization of DNA Extraction and PCR Amplification Protocols Using the Model Plant Arabidopsis thaliana
Abstract
Arabidopsis thaliana, a small spring annual plant is used as a model in plant genetics and biotechnology due to some advantages like relatively rapid life cycle (8 weeks from germination to production of seeds) and small genome size. Because of these advantages series of loss of function and gain of function mutant plants were generated using ecotypes of this plant in order to research involvement of proteins and phytohormones in plants. The most commune approach to test the obtained mutants for their homozygosis in response to the T- DNA (transfer DNA) insert is by the use of the Polymerase Chain Reaction (PCR) method. There were always problems regarding the isolation and amplification of DNA mentioned in the specific literature this was the main reason why this research was focused on comparing two extraction protocols and two PCR master mixes. The results obtained were satisfying leading us to the conclusion that both extraction methods and amplification master mixes can be implemented further into the research and maybe be also transferred to species phylogenetically related with the model plant.
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