Fig 1.

Lack of strand specificity in detecting negative-sense RNA due to false priming during cDNA synthesis: 10⁶ (lanes 1 to 9) and 10³ (lanes 10 to 18) HEVT1FG RNA copies were reverse transcribed in the presence of AMV-RT (lanes 1, 2, 3, 10, 11, and 12), Superscript RT-III (lanes 4, 5, 6, 13, 14, and 15), or no enzyme (lanes 7, 8, 9, 16, 17, and 18). Lanes 1, 4, 7, 10, 13, and 16 represent RT reactions with primer HEVF1, lanes 2, 5, 8, 11, 14, and 17 represent RT reactions with primer HEVR1, and lanes 3, 6, 9, 12, 15, and 18 represent RT reactions with no primer. A 10-μl volume of cDNA was used for the first-round PCR, and 10 μl of the first PCR product was used for the nested PCR.