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Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood

reactions. The expected amplification products for each chromosome were obtained, indicating that the genomic DNA isolated using the DNA Stat kit contains a complete representation of all human chromosomes (data not shown).

Isolated Genomic DNA for Reference Laboratory Analysis

The DNA Stat kit was used in a reference laboratory setting (Cenetron, Inc.) to isolate genomic DNA from patient whole blood samples. The resulting DNA was used as template for quantitative PCR analysis with primers specific to Factor V. Results from PCR reactions using 5 ml of template DNA isolated from six individual whole blood samples showed that each reaction produced the expected 107-bp PCR product and, therefore, the isolated genomic DNA is suitable for use in clinical reference laboratory assays (data not shown).

Conclusions

The quick and easy-to-use DNA Stat is the choice for isolating PCR-ready genomic DNA from whole blood. The isolated DNA is high molecular weight (40 kb) and an excellent template for PCR analysis. Because of the kits unique spin-cup design, there is no need to remove RBC or pellet WBC before beginning the procedure. Additionally, there is no need for alcohol precipitation, no need to digest with proteases, and no need to alter the protocol with a 25-l to 200-l blood volume. Furthermore, all incubations are carried out at room temperature, all steps are performed in a single microspin cup, and all isolations are easily completed in 40 minutes. An aliquot of 1 ml to 5 ml of the isolated genomic DNA is sufficient for most PCR analyses including clinical reference laboratory assays.

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