Comparison of different collection procedures and two methods for DNA isolation from saliva

Jaroslava Durdiaková, Natália Kamodyová 1 , Daniela Ostatníková 2 , Barbora Vlková 1  and Peter Celec
  • 1 Institute of Molecular Biomedicine, Comenius University, Bratislava, Slovakia
  • 2 Institute of Physiology, Comenius University, Bratislava, Slovakia
  • 3 Institute of Pathophysiology, Comenius University, Bratislava, Slovakia
  • 4 Department of Molecular Biology, Comenius University, Bratislava, Slovakia

Abstract

Background: The non-invasive, flexible and easy sample collection makes saliva an interesting source of DNA for research and diagnostic purposes. The aim of our study was to find the most suitable collection method for biological material from the oral cavity and the most effective DNA isolation technique for further analytic applications.

Methods: DNA was isolated from swabs, Salivette saliva, whole saliva and samples collected with a commercial set for scraping of buccal cells. Phenol-chloroform extraction and isolation using a silica membrane based commercial kit were compared. Quantity of bacterial and human genomic DNA was estimated using real time PCR. The effects of storage conditions on DNA recovery were assessed.

Results: Sample collection techniques significantly affected the quantity of DNA for both, silica membrane based and phenol-chloroform isolations. Whole saliva provided the largest number of bacterial and human genome copies after both extraction methods. Storage for 36 months at –20°C reduced recovery of human genomic DNA five times after silica membrane based extraction and 10 times after phenol-chloroform isolation.

Conclusions: Whole saliva was found to be the most suitable material for human and bacterial DNA isolation. Both compared methods are useful considering the quantity of extracted DNA.

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Clinical Chemistry and Laboratory Medicine ( CCLM) publishes articles on novel teaching and training methods applicable to laboratory medicine. CCLM welcomes contributions on the progress in fundamental and applied research and cutting-edge clinical laboratory medicine. It is one of the leading journals in the field, with an impact factor of over three. CCLM is the official journal of nine national clinical societies and associated with EFLM.

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