Published in Scientific Bulletin. Series F. Biotechnologies, Vol. XXVIII, Issue 2
Written by Claudia DÎRLĂ, Maria Rodica GURĂU, Fănel OȚELEA, Nicoleta DRĂGUȚ, Georgeta ȘTEFAN, Marius Andrei MĂNESCU, Doina DANEȘ
Currently, the ASF (African Swine Fever) diagnosis is carried out by the detection of viral DNA. The successful amplification of the targeted DNA fragment needs a proper quantity of the genetic material. The aim of this study was to compare the yield and quality of DNA extracted using two dedicated commercial kits. It was compared the product obtained using the Pure Link Genomic DNA Mini Kit (Invitrogen) and QIAamp Cador Pathogen Mini Kit (Qiagen). The DNA has been quantified using Qubit DNA HS Assay Kit (Qubit 3.0 Fluorometer, ThermoFisher Scientific), a highly selective over RNA and accurate tool for DNA at levels between 10 pg/µL and 100 ng/µL. The standard DNA of the ASF virus has been diluted 10-2, 103, 10-4, 10-5 and 10-6 as both extracts obtained with Pure Link Genomic DNA Mini Kit (Invitrogen) and the QIAamp Cador Pathogen Mini Kit (Qiagen). According to the dilutions (10-2 to 10-6), the DNA yield with the QIAamp Cador Pathogen Mini Kit was 132 ng/µl, 71 ng/µl, 45 ng/µl, 21.2 ng/µl and 10.2 ng/µl, respectively. The yield obtained with the Pure Link Genomic DNA Mini Kit was 126 ng/µl, 65 ng/µl, 31 ng/µl, 15.2 ng/µl and 4.2 ng/µl, respectively. The highest concentration was obtained using the QIAamp Cador Pathogen Mini Kit (Qiagen). In conclusion, the sensitivity of the Qiagen Kit is more suitable to be used for further investigation on the ASF genome.
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