Open Access Highly Accessed Research article

Critical evaluation of HPV16 gene copy number quantification by SYBR green PCR

Ian Roberts1*, Grace Ng1, Nicola Foster1, Margaret Stanley2, Michael T Herdman1, Mark R Pett1, Andrew Teschendorff3 and Nicholas Coleman12

Author Affiliations

1 MRC Cancer Cell Unit, Hutchison/MRC Research Centre, Hills Road, Cambridge. CB2 0XZ, UK

2 Department of Pathology, Cambridge University, Tennis Court Road, Cambridge. CB2 1QP, UK

3 Cancer Research UK, Li Ka Shing Centre, Cambridge Research Institute, University of Cambridge, CB2 0RE, UK

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BMC Biotechnology 2008, 8:57  doi:10.1186/1472-6750-8-57

Published: 24 July 2008

Additional files

Additional file 1:

Crossing point values used in critical evaluation of HPV16 gene copy number quantification by SYBR green PCR. The Microsoft Excel workbook of Additional file 1 contains four worksheets. 1: NA6 Standard Curves. The crossing point values used in generation of external calibration curves for absolute quantification of viral E2, viral E6 and host HMBS genes are presented. For each gene, four runs of a seven point NA6 titration series were conducted in triplicate. 2: Accuracy Test. The crossing point values used to determine viral load and physical state of a three point serial dilution of a mixture of test gDNA and HPV16 plasmid DNA are presented. Three runs were undertaken at each titration point in triplicate. 3: Assessment of Cell Lines. The crossing point values used to derive viral load and physical state of five cervical carcinoma cell lines are presented. Each cell line was assessed in three separate runs, and each reaction was performed in triplicate. 4: Assessment of Clinical Samples. The crossing point values used to derive viral load and physical state of 14 squamous cell cervical carcinoma samples are presented. One run was undertaken for each sample, and all reactions were performed in triplicate.

Format: XLS Size: 94KB Download file

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