PCR for DNA amplification
SEPARATION, AMPLIFICATION, POSITIVE
PCR was performed using a set of specific primers proposed by Sitnik et al. (2004) for HBV and Shahinsaz et al. (Shahinsaz et al., 2006) for HDV, the characteristics of which presented in Tables S1 and S2 [In the absence of a PCR product of a given size, a second round of PCR was performed with nested primers and under the same conditions, instead of DNA, PCR products of the first round were used. Positive and negative control samples were used during PCR. Both rounds of PCR were carried out with a total volume of the reaction mixture of 50 µl.Amplification of PCR products was carried out on a GeneAmp® PCR System 9700 thermal cycler (Applied Biosystems™, USA). Detection and separation of PCR products was carried out in 2% agarose gel, on TAE buffer, using GeneRuler 100 bps DNA Ladder (Thermo Fisher Scientific™, USA). Gels were stained using UltraPure™ Ethidium Bromide (Invitrogen™, USA). Samples with specific PCR products were further used for purification and subsequent sequencing.