Figure 6

5'-VVVVVVVVAA-3' Primers Enable Random Multiplex Amplification of Viral RNA from Human Plasma Inoculated with Coxsackie Virus A7. A. 1 ml of human plasma was inoculated with 5 μl of suspended Coxsackie Virus A7 (ATCC), filtered and incubated for 2 hours with nucleases. Remaining nucleic acids were purified with the QiaAmp UltrasSens Virus Kit (Qiagen) and subjected to first strand cDNA synthesis using either Omniscript or Superscript reverse transcriptase. Virus-specific primers were used to amplify either a 200 bp or a 1.4 Kb portion of the viral genome. B. The same samples were then subjected to random multiplex PCR with 5'-VVVVVVVVAA-5' primers as detailed in the Methods section. Amplicons were then visualized on an ethidium bromide impregnated agarose gel.