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Oragene and Whole Genome Amplification with GenomiPhi

J. Chartier
DNA Genotek Inc., Ottawa, ON, Canada

DNA collected with Oragene is successfully amplified using the GenomiPhi DNA Amplification Kit and has the potential to generate near-unlimited quantities of DNA for downstream experiments.


Introduction
As the genotyping of single nucleotide polymorphisms (SNPs) becomes more automated and less costly, the limiting factor for genome-wide genetic analysis is becoming the limited amount of DNA that can be collected from study subjects. A renewable source of genomic DNA is desirable. The traditional method of transforming lymphoblastic cell lines is usually too expensive for population studies with large sample sizes. A less expensive alternative is whole genome amplification (WGA), which allows in vitro production of numerous copies of the entire genome.

Multiple displacement amplification (MDA) is a WGA method that uses Phi29 DNA polymerase and random priming. MDA generates high-fi delity genomic copies with minimal bias (ref. 1).

The GenomiPhi DNA Amplification Kit from Amersham Biosciences utilizes MDA to exponentially amplify genomic DNA. The purpose of this study was to determine if DNA collected with Oragene could be amplified with GenomiPhi and to test the quality of the amplified DNA for PCR and SNP genotyping.


Materials and Methods
DNA collection
Saliva was collected from 10 donors using Oragene DNA Self-Collection Kits. DNA was purified from 200 L aliquots of Oragene/saliva samples using the ethanol precipitation protocol supplied with the kits. Purifi ed DNA was redissolved in 200 L of TE buffer (10 mM Tris-HCl, 1 mM EDTA,
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