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The Importance of Careful Blood Processing in Isolation of Cell‐Free DNA
Author(s) -
PAGE KAREN,
POWLES TOM,
SLADE MARTIN J.,
DE BELLA MANUELA TAMBURO,
WALKER ROSEMARY A.,
COOMBES R. CHARLES,
SHAW JACQUELINE A.
Publication year - 2006
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1368.042
Subject(s) - dna , centrifugation , amplicon , dna extraction , cell free fetal dna , whole blood , cell , differential centrifugation , extraction (chemistry) , microbiology and biotechnology , biology , chemistry , chromatography , polymerase chain reaction , biochemistry , genetics , gene , immunology , pregnancy , fetus , prenatal diagnosis
In healthy individuals, the source of cell‐free plasma DNA is predominantly apoptotic, whereas, increased plasma DNA integrity is seen in cancer patients. Therefore, it is important to carefully isolate absolutely “cell‐free” plasma DNA. Plasma DNA from 30 healthy females was analyzed using 4 PCR amplicons of increasing size, comparing standard blood processing with additional centrifugation steps prior to DNA extraction. Cellular DNA contamination, indicated by positive amplicons >300 bp was eliminated only after the extra centrifugation step. This highlights the importance of careful processing in preparation of cell‐free plasma DNA as a tool for cancer detection and we recommend the use of a microcentrifuge spin, prior to DNA extraction.