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Fig. 2 | Clinical Epigenetics

Fig. 2

From: Comprehensive evaluation of targeted multiplex bisulphite PCR sequencing for validation of DNA methylation biomarker panels

Fig. 2

Pre-sequencing optimisation of multiplex PCR primers. a PCR products of singleplex amplification of 33 individual primer pairs from the breast cancer panels run on 2% agarose gel. The gels show the specificity of all the primer pairs and PCR products of the correct size (100–130 bp) with minimal primer dimer formation. (−) no template; (+) bisulphite-treated test DNA template (10 ng); (L) 100 bp DNA Ladder. b Singleplex primers were pooled into their respective multiplex panels, and the outcome of multiplex PCR reactions is shown at different primer concentrations (20 μM, 10 μM, 5 μM and 1 μM) and at different annealing temperatures (55 °C, 56 °C and 57 °C). c PCR products from the multiplex panels testing DNA input amounts of 10 ng, 5 ng, 2.5 ng, 1.25 ng and 0.625 ng of bisulphite-treated control DNA; (+) test DNA; (−) no template control

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