Key result
The in vial RNA-PCR method, which performs reverse transcription and PCR sequentially in the same vial using UNG and dUTP separated by a wax barrier, successfully produced UNG-sensitive amplicons to prevent carry-over contamination.
Population
HIV-1 RNA isolated from H9 cells infected with the DIB strain of HIV-1
Comparison
In vial RNA-PCR technique using… vs Reverse transcription and PCR both performed…
Design
Preclinical
Authors
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May reduce carry-over contamination in RNA-PCR; extends UNG-dUTP approach but leaves open clinical translation.
The in vial RNA-PCR (IVRP) method successfully extends the UNG-dUTP decontamination approach to RNA-PCR by physically separating RT and PCR steps with a wax barrier.
Udaykumar et al. (1993) studied Carry-over contamination in RNA-PCR. In vial RNA-PCR (IVRP) with UNG and dUTP vs. Conventional RT-PCR with dTTP was evaluated on UNG-sensitivity of amplified products. The in vial RNA-PCR method, which performs reverse transcription and PCR sequentially in the same vial using UNG and dUTP separated by a wax barrier, successfully produced UNG-sensitive amplicons to prevent carry-over contamination.
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