VALIDATION AND RAPID EXTRACTION OF NUCLEIC-ACIDS FROM ALCOHOL-PRESERVED TICKS

A method has been developed to extract DNA from alcohol-preserved tick s (Acari: Ixodidae and Argasidae). The method combines the lysing prop erty of the chaotropic agent guanidinium thiocyanate (GuSCN) and the n ucleic acid-binding property of diatomaceous earth (fossilized cell wa lls of unicellular algae). Debris from the tick is removed in several sequential washing steps. To monitor the efficiency of this method, a polymerase chain reaction (PCR) was designed to amplify the 16S mt rRN A gene of five tick genera (Dermacentor Fabricius, Haemaphysalis Koch, Rhipicephalus Koch, Argas Latreille and Ixodes Latreille). Detection of amplification products from this PCR indicated that DNA had been su ccessfully extracted and that Taq-polymerase inhibitors were absent. T he extraction method, therefore, enables purification of DNA such that enzymatic analysis is possible.