Real-time PCR reactions were performed using Roche Lightcycler TaqMan master mix. Each reaction consisted of 1X Roche Lightcycler TaqMan master mix (Roche Diagnostics, Indianapolis, Indiana, USA), HPV-specific primer pairs and fluorescently-tagged probes for types 6, 11, 16, and 18 in a total reaction volume of 10 ml as previously described [15 (link)]. Real-time PCR reactions that target the E1 region of HPV were performed using Roche Lightcycler Sybr Green master mix (Roche Diagnostics), and sequencing of PCR products was performed as previously described [15 (link)].
Protocol detail
Real-time PCR for HPV detection
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Diagnostics
Primer
Sybr green
Corresponding Organization : University of California, San Francisco
Other organizations : Cancer Research And Biostatistics, University of North Carolina at Chapel Hill, University of California, Los Angeles, Rutgers, The State University of New Jersey
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Variable analysis
independent variables
- HPV-specific primer pairs and fluorescently-tagged probes for types 6, 11, 16, and 18
- Expression levels of HPV types 6, 11, 16, and 18
- Roche Lightcycler TaqMan master mix
- Roche Lightcycler Sybr Green master mix
- Total reaction volume of 10 ml
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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