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Automated genomic DNA extraction from saliva using the QIAxtractor
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Henry Keijzer
Published/Copyright:
March 11, 2010
Abstract
Background: Venipuncture is an invasive procedure to obtain whole blood in order to obtain high quality and sufficient amounts of genomic DNA. Obtaining DNA from non-invasive sources is preferred by patients, medical doctors and researchers. Saliva collected with cotton swabs (Salivette®) is increasingly being used to study chemical compounds, and it can also be a source of DNA. However, extracting DNA from Salivettes is very laborious and time consuming. Therefore, we developed a protocol for automated genomic DNA extraction from saliva collected in Salivette® using the QIAxtractor.
Methods: Saliva (0.1–2.0 mL) was collected by chewing on a Salivette® for 1–2 min. A total of 70 samples, collected from healthy volunteers, were extracted with the QIAxtractor robot and a Qiagen DX reagent pack. Quantity and quality was assessed using UV spectrometry and real-time polymerase chain reaction (PCR) (substitution at position –729 in the CYP1A2 gene).
Results: The average DNA concentration from the saliva samples was 6.0 μg/mL (95% CI 5.4–6.6 μg/mL). In 100% of the saliva samples, PCR products were detected with an average cycle threshold of 23.1 (95% CI 22.6–23.6).
Conclusions: DNA can be extracted in sufficient amounts from Salivette® with a fully automated system with a short turnaround time. Real-time PCR can be performed with these samples.
Clin Chem Lab Med 2010;48:641–3.
Received: 2009-10-19
Accepted: 2010-1-12
Published Online: 2010-03-11
Published in Print: 2010-05-01
©2010 by Walter de Gruyter Berlin New York
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