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A high-throughput protocol for isolating cell-free circulating tumor DNA from peripheral blood.
A high-throughput protocol for isolating cell-free circulating tumor DNA from peripheral blood.
- Source :
-
BioTechniques [Biotechniques] 2019 Feb; Vol. 66 (2), pp. 85-92. - Publication Year :
- 2019
-
Abstract
- The analysis of cell-free circulating tumor DNA (ctDNA) is potentially a less invasive, more dynamic assessment of cancer progression and treatment response than characterizing solid tumor biopsies. Standard isolation methods require separation of plasma by centrifugation, a time-consuming step that complicates automation. To address these limitations, we present an automatable magnetic bead-based ctDNA isolation method that eliminates centrifugation to purify ctDNA directly from peripheral blood (PB). To develop and test our method, ctDNA from cancer patients was purified from PB and plasma. We found that allelic fractions of somatic single-nucleotide variants from target gene capture libraries were comparable, indicating that the PB ctDNA purification method may be a suitable replacement for the plasma-based protocols currently in use.
- Subjects :
- Biomarkers, Tumor blood
Biomarkers, Tumor isolation & purification
Cell-Free Nucleic Acids isolation & purification
Circulating Tumor DNA isolation & purification
High-Throughput Nucleotide Sequencing
Humans
Mutation
Neoplasms genetics
Cell-Free Nucleic Acids blood
Circulating Tumor DNA blood
High-Throughput Screening Assays methods
Neoplasms blood
Subjects
Details
- Language :
- English
- ISSN :
- 1940-9818
- Volume :
- 66
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- BioTechniques
- Publication Type :
- Academic Journal
- Accession number :
- 30744412
- Full Text :
- https://doi.org/10.2144/btn-2018-0148