The quality and quantity of DNA extracted from the tumour cells is the key to successful downstream analysis.
- A poor DNA extraction method can lead to increased assay failure and increase the risk of false positive and false negative results
DNA extraction
• Extraction methods need to be robust, reliable and yield as much DNA as possible• Many DNA extraction methods are available, including kit-based methods, eg QIAamp® DNA minikit (Qiagen)
(Eberhard 2008)
DNA quantification
DNA quantification is optional, but may be useful for prioritising exons for screening and troubleshooting assay failure
- Quantitative PCR (Q-PCR) is more accurate than spectrophotometry
- Q-PCR measures only the amplifiable DNA in a sample so it is also a measure of DNA quality. DNA that has broken down during storage is therefore accurately quantified