Fast DNA Purification For SNP Analysis

DNA Stat^™ blood kit ideal for SNP analysis

Fast DNA Purification for SNP Analysis

Sylvia Norman •  Gothami Padmabandu • Jeff Braman

Single nucleotide polymorphism (SNP) genotyping is a valuable tool for identifying genetic mutations associated with predisposition to complex diseases and traits such as cancer, cardiovascular disease, diabetes, and obesity. The success of SNP analyses lies in isolating highly purified, PCR-quality genomic DNA from whole blood samples.

The DNA Stat^™ blood kit¹ offers a fast and easy way to isolate PCR-quality genomic DNA from whole blood samples ranging in volume from 25 µl to 200 µl. To demonstrate the utility of the DNA Stat blood kit for SNP allelic discrimination assays, we isolated genomic DNA from 15 individual whole blood samples using 100 ml whole blood for each preparation. The resulting genomic DNA was then used as a template in SDF-1(3' A) allelic discrimination assays. The SDF-1(3' A) allelic discrimination kit² detects a single nucleotide mutation which can predispose an HIV-1 infected individual to either delayed or accelerated onset of AIDS. This kit includes a mixture of two SDF-1 allele-specific molecular beacons with an exact sequence match to each of the two target sequence variants. Each of the molecular beacons is labeled with a fluorophore making it possible to detect all three allelic combinations: TET fluorescence indicates wild-type homozygosity, FAM fluorescence indicates mutant allele homozygosity and TET/FAM fluorescence indicates heterozygosity.

The results (Figure 1) demonstrate that DNA Stat Blood Kit genomic DNA preparations are ideal for use in allelic discrimination assays. All three possible genotypes were detected in the 15 individual samples assayed: two mutant homozygotes, four heterozygotes and nine wild-type homozygotes (each data point represents duplicate samples). Two microliters of a 50 µl DNA sample preparation were used as template for each PCR reaction. Therefore, each DNA sample preparation provides sufficient template for 25 allelic discrimination assays.

The fast and easy isolation method makes the kit ideal for not only SNP genotyping, but for other allelic discrimination assays as well. 

REFERENCES

1. Norman, S. and Braman, J. (2000) Strategies 13: 154-156.
2. Hu, X. and Lingyu, C. (2000) Strategies 13: 24-26.
3. Horner, R. et al, (2001) Strategies 14: 1-4