Open Access Research article

A closed-tube methylation-sensitive high resolution melting assay (MS-HRMA) for the semi-quantitative determination of CST6 promoter methylation in clinical samples

Lampros Dimitrakopoulos1, Panagiotis A Vorkas13, Vasilis Georgoulias2 and Evi S Lianidou1*

Author Affiliations

1 Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens, 15771, Greece

2 Department of Medical Oncology, University General Hospital of Heraklion, PO BOX 1352, Crete, 71110, Greece

3 Present address: Biomolecular Medicine, Department of Surgery and Cancer, Imperial College London, London, SW7 2AZ, UK

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BMC Cancer 2012, 12:486  doi:10.1186/1471-2407-12-486

Published: 22 October 2012

Additional files

Additional file 1:

Figure S1. Optimization of the annealing temperature of the MS-HRMA assay for CST6 promoter methylation. Normalized melting curves and first derivative plots for a) 63ºC: Black: 0%, red: 1%, blue: 10%, green: 50%, yellow: 100% methylation b) 61ºC: Black: 0%, red: 1%, blue: 10%, yellow: 50%, green: 100% methylation and c) 60ºC: Black: 0%, red: 1%, blue: 10%, green: 50%, yellow: 100% methylation.

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Additional file 2:

Figure S2. Specificity of MS-HRMA assay for CST6 promoter methylation: PCR products of the SB modified positive controls and genomic DNA (unconverted). 1) DNA ladder 2) negative control (H2O), 3) 0% methylated control 4) 1% methylated control 5) 10% methylated control 6) 50% methylated control 7) 100% methylated control 8) genomic DNA (unconverted).

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Additional file 3:

Table S1. CST6 methylation status in 10 paired breast cancer and 10 adjacent non-cancerous tissues as evaluated by both the developed MS-HRMA and MSP [26] assays.

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