Cell-Free Circulating DNA
Purification and Analysis Techniques
Circulating cell-free DNA is poised to transform cancer diagnosis and care; however, it carries technical challenges such as low abundance, fragmentation and sensitivity to biospecimen handling. Development of clinically useful assays hinges on understanding the unique technical aspects of working with cell-free DNA as a substrate. This book provides an in-depth summary of the technical issues impacting cell-free DNA purification from blood plasma and analysis in a cancer context, including design of PCR assays, sequencing library preparation and methylation analysis. Emerging fields such as extracellular vesicles and blood nucleases are also covered, as well as basic biology and considerations pertinent to biobanking.
- Biology of Circulating DNA in Health and Disease (Ahuva Odenheimer-Bergman, Havell Markus and Muhammed Murtaza)
- Preanalytics and Quality Control of Different Substrates for Circulating DNA Studies (Fay Betsou and Wim Ammerlaan)
- Considerations in Biobanking Blood Samples and Plasma Volume for Circulating DNA Studies (Kate Mahon, Goli Samimi and Kristina Warton)
- Maximizing Yield from Plasma Circulating DNA Extraction (Florence Mauger and Jean-François Deleuze)
- Optimal Design of PCR Assays for Circulating DNA (Rikke F Andersen)
- Preparation of Next-Generation Sequencing Libraries for Sequencing Circulating DNA (Katrin Heider, Florent Mouliere and Christopher G Smith)
- Blood Nucleases Affecting Circulating DNA in Serum and Plasma (Gustavo Barra)
- Isolating Circulating Exosomes as Biomarkers: Challenges and Opportunities (Alexander Semaan and Anirban Maitra)
- DNA Methylation Analysis of Circulating DNA Biomarkers (Kristina Warton, Clare Stirzaker, Goli Samimi and Susan Clark)
Readership: Graduate students and researchers in Cancer biology, Oncology and Biomedical Sciences.
- The book systematically examines all steps of handling and analysing circulating DNA, from blood collection though DNA analysis, including parameters that are important for successful commercialisation of resulting diagnostic tests
- Chapters are written by researchers with a specific interest and practical experience in the quality control of each step
- While published academic studies often focus on the biological and clinical significance of data, the main theme of this text is the technical considerations of working with circulating DNA, which are often omitted from the literature. The problems stemming from technical inconsistencies have been recognised in a number of high impact papers, and this book brings together international expertise on the topic