Samsonova Jeanne V, Saushkin Nikolay Yu, Voronkova Valery N, Stolpovsky Yuri A, Piskunov Aleksei K
Faculty of Chemistry, Lomonosov Moscow State University, Moscow, 119991, Russia.
Vavilov Institute of General Genetics, Moscow, 119991, Russia.
Biochem Genet. 2024 Jul 12. doi: 10.1007/s10528-024-10882-7.
While dried blood spots are a convenient source of genetic material, they are usually associated with a lower DNA yield than from a native sample. The study evaluated the DNA yield from dried blood samples prepared on glass fibre and cellulose membranes and investigated the reasons for the yield reduction. The extraction of total DNA from membrane-dried blood samples was optimized by spin-column extraction method. It was shown that preliminary short-term (20 min) solubilization of a dried matrix in an aqueous medium, followed by standard extraction protocols for the mixture of the eluate with membranes, provides the highest DNA yield. The yield of DNA from a glass fibre membrane was 40-50% lower compared to a native sample, but on average, two times higher than from a conventional cellulose membrane (filter paper). The reduction of DNA yield when using a dried sample was found to be due to partial retention of nucleic acids by the membrane material during the lysis stage.
虽然干血斑是一种方便的遗传物质来源，但与天然样本相比，它们通常产生的DNA产量较低。该研究评估了在玻璃纤维和纤维素膜上制备的干血样本的DNA产量，并调查了产量降低的原因。通过旋转柱提取法优化了从膜干血样本中提取总DNA的方法。结果表明，将干燥的基质在水性介质中进行初步短期（20分钟）溶解，然后对洗脱液与膜的混合物采用标准提取方案，可提供最高的DNA产量。与天然样本相比，玻璃纤维膜的DNA产量低40 - 50%，但平均而言，比传统纤维素膜（滤纸）高出两倍。研究发现，使用干样本时DNA产量降低是由于在裂解阶段核酸被膜材料部分保留所致。
