Whole-genome amplified DNA from stored dried blood spots is reliable in high resolution melting curve and sequencing analysis

We wanted to compare melting curves and sequencing results from wgaDNA derived from DBS samples with gDNA derived from whole blood.gDNA was extracted from whole blood obtained from 10 patients with lone atrial fibrillation (mean age 22.3 years). From their newborn DBS samples, stored at -24°C, genomic DNA was extracted and whole-genome amplified in triplicates. Using high resolution melting curve analysis and direct sequencing in both wgaDNA and gDNA samples, all coding regions and adjacent intron regions of the genes SCN5A and KCNA5 were investigated.Altered melting curves was present in 85 of wgaDNA samples and 81 of gDNA samples. Sequence analysis identified a total of 31 variants in the 10 wgaDNA samples. The same 31 variants were found in the exact same pattern of samples in the gDNA group. There was no false positive or negative sequence variation in the wgaDNA group.The use of DNA amplified in triplicates from DBS samples is reliable and can be used both for high resolution curve melting analysis as well as direct sequence analysis. DBS samples therefore can serve as an alternative to whole blood in sequence analysis.The challenge to investigate the genetic basis of inherited diseases requires large amount of DNA preferably obtained from drawn blood samples. However, in some cases drawn blood is not available. Genomic DNA (gDNA) from dried blood spots (DBS) samples has previously been shown to be reliable for genetic testing [1], but the amount of gDNA is limited and as such comprehensive investigations including new candidate gene screening in these patients might be impossible. In those situations, amplification of small amounts of gDNA using whole-genome amplification (WGA) can be beneficial. WGA is a method based on the multiple displacement amplification technology [2]. The technology replicates-using a DNA polymerase-up to 100 kb without dissociating from the gDNA template. The polymerase moves along the DNA template strand displacing the complementary