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Effect of the DNA isolation method on the results of multiple fluorescent PCR and DNA fragment analysis
Authors:Nagy Bálint  Bán Zoltán  Papp Zoltán
Affiliation:Semmelweis Egyetem, Altalános Orvostudományi Kar, I. Szülészeti es Nogyógyászati Klinika, Budapest.
Abstract:INTRODUCTION: The quality of the nucleic acids has tremendous effects on the results of molecular diagnostic tests. METHODS: The authors have isolated DNA from amniotic fluid samples by using rezin-binding and silica adsorption methods. In the case of the rezin-binding technique they used both frozen and fresh samples. The quality of the isolated DNA was compared by fluorescent PCR and DNA fragment analysis. RESULTS: With the use of the rezin-binding method on frozen samples the authors obtained more PCR fragments following the fluorescent PCR amplification. The gender and the aneuploidy screening is easier from the electrophoretogramms using better quality DNA. Out of the three studied DNA isolation method the silica adsorption seemed to be the best for the fluorescent-PCR and DNA fragment analysis. DISCUSSION: The authors found the silica adsorption method is more suitable for DNA isolation from amniotic fluids for fluorescent PCR and DNA fragment analysis amplification. They suppose silica adsorption DNA isolation results in a total removal of the PCR inhibitors.
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