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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.

Authors :
Pandoh PK
Corbett RD
McDonald H
Alcaide M
Kirk H
Trinh E
Haile S
MacLeod T
Smailus D
Bilobram S
Mungall AJ
Ma Y
Moore RA
Coope R
Zhao Y
Jones SJ
Holt RA
Karsan A
Morin RD
Marra MA
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.

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