Automated genomic DNA extraction from saliva using the QIAxtractor.
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 microg/mL (95% CI 5.4-6.6 microg/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.
Keijzer
Henry
H
Department of Clinical Chemistry and Hematology, Gelderse Vallei Hospital, Ede, The Netherlands. keijzerh@zgv.nl
Endenburg
Silvia C
SC
Smits
Marcel G
MG
Koopmann
Miriam
M
D001331:Automation; D004247:DNA / Q000302:isolation & purification*; D015894:Genome, Human; D006801:Humans; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D012463:Saliva / Q000737:chemistry*
D011933:Reagent Kits, Diagnostic; D004247:DNA