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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
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