The importance of careful blood processing in isolation of cell-free DNA.
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.
Page
Karen
K
Department of Cancer Studies and Molecular Medicine, Robert Kilpatrick Clinical Sciences Building, University of Leicester, Leicester, LE2 7LX, United Kingdom. kd18@leicester.ac.uk
Powles
Tom
T
Slade
Martin J
MJ
DE Bella
Manuela Tamburo
MT
Walker
Rosemary A
RA
Coombes
R Charles
RC
Shaw
Jacqueline A
JA
D001774:Blood Chemical Analysis; D002498:Centrifugation; D004247:DNA / Q000097:blood / Q000302:isolation & purification; D005260:Female; D006801:Humans; D009369:Neoplasms / Q000235:genetics; D010949:Plasma / Q000737:chemistry; D016133:Polymerase Chain Reaction
D004247:DNA