( isolations were performed under identic...)Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood

isolations were performed under identical conditions using the genomic DNA isolation procedure described in the kits manual (Figure 2). PCR analysis was performed using 1 ml of the isolated genomic DNA as template and primers specific for 1-antitrypsin (a1-AT). The results demonstrate that 1 ml of genomic DNA template isolated from a range of blood volumes is sufficient for amplification of the expected 980-bp PCR product.

Amplification of Various PCR Target Sizes

Fig.3

Genomic DNA isolated from blood using the DNA Stat kit is high molecular weight (approximately 40 kb in size) (Figure 3). To further test the integrity of the isolated genomic DNA, 1-ml aliquots of a 50-ml prep were used as templates for amplifying a range of PCR target sizes. We used primers specific to aldolase B (Aldo: 250 bp), T-cell receptor b-chain gene (TCRb: 500 bp), and a1-antitrypsin (a1-AT: two primer sets amplifying 980-bp and 5.2K-bp PCR targets) in PCR reactions with 1 ml al of genomic DNA. Genomic DNA isolated using the DNA Stat kit is suitable for amplifying PCR targets ranging in size from 250 bp to 5.2 kb (Figure 4).

Fig.4

We also assessed genomic DNA quality by amplifying PCR targets on each of the human chromosomes. Primers designed to amplify targets containing simple sequence repeats on chromosomes 1 through 22 plus X and Y were used for this analysis (Human MapPairs primers, Research Genetics: TCRb primers were used for chromosome 7). The resulting PCR products ranged in size from 202 bp to 500 bp. From 100 ml of whole blood, 1 ml of DNA was used as a template for PCR
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