Figure 1.
Figure 1. Real-time strand-specific quantitative assay for the detection of HCV RNA–negative and –positive strands. Reverse transcription was done at 70° C using thermostable enzyme Tth and the first round of PCR amplification was limited to 20 cycles. The nested round employed LightCycler FastStart DNA Master SYBR Green I (Roche Diagnostics). (A) Detection of negative-strand HCV RNA synthetic template. (B) Detection of positive-strand HCV RNA synthetic template. (C) Specificity of the assay for the detection of negative-strand HCV RNA: reverse transcription was done with positive-sense primer using serial dilution of positive-strand HCV RNA as template. (D) Nonnested real-time PCR detection of HCV RNA–positive strand using MMLV for the reverse transcription step.

Real-time strand-specific quantitative assay for the detection of HCV RNA–negative and –positive strands. Reverse transcription was done at 70° C using thermostable enzyme Tth and the first round of PCR amplification was limited to 20 cycles. The nested round employed LightCycler FastStart DNA Master SYBR Green I (Roche Diagnostics). (A) Detection of negative-strand HCV RNA synthetic template. (B) Detection of positive-strand HCV RNA synthetic template. (C) Specificity of the assay for the detection of negative-strand HCV RNA: reverse transcription was done with positive-sense primer using serial dilution of positive-strand HCV RNA as template. (D) Nonnested real-time PCR detection of HCV RNA–positive strand using MMLV for the reverse transcription step.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal