In order to facilitate experimental optimization, we defined a standard MDA protocol with a reaction volume of 40 μL and consisting of phi29 DNA polymerase and its corresponding buffer, 1 mM of dNTPs, and 2 μM or 50 μM of random exonuclease-resistant primers. A recombinant 12-kb plasmid (HCV plasmid) containing a 9-kb HCV genotype 1a genome from our previous study (20 (link)) was used as a positive control. The reaction was incubated at 30°C for 16 h, followed by heat-inactivation at 65°C for 10 min. In addition to visualization on a 0.8% agarose gel, MDA yield was quantitated by the Quant-iT PicoGreen dsDNA Assay kit (QIAGEN, Valencia, CA) after purification using the QIAamp DNA mini kit. Due to the extreme sensitivity of the techniques (5 (link)), all MDA experiments were performed in strict adherence to the recommendations of Champlot et al. to avoid possible contamination (21 (link)).