Circulating DNA is fragments of DNA which can be found in the blood of healthy
as well as diseased individuals. Higher levels of these nucleic acid molecules
can be found in diseased and pregnant individuals in contrast to healthy controls.
The origin of circulating DNA has not been elucidated, but release of DNA after
apoptosis or necrosis or active release by living cells has been hypothesized. It
was concluded in this study that apoptosis or necrosis may only be a minor
source of circulating DNA and that release of DNA by living cells might play a
major role in the origin, while disturbance of the equilibrium between release by
living cells and clearance mechanisms may cause the rise in the levels of
circulating DNA observed in different conditions.
Before circulating DNA can be analyzed, it has to be isolated from the blood. A
number of different preanalytical conditions can have an impact on the quantity
and quality of circulating DNA that can be isolated. Furthermore, the choice of
isolation and quantification method may also influence the results obtained.
Quantitative analysis of circulating DNA was done by real-time PCR analysis of
the &Globin gene and the DNA levels obtained for healthy controls and cancer
patients correlated with levels reported in the literature.
Characterization of total circulating DNA may be beneficial in diagnosis and
prognosis and may also contribute to determining the source and function of
circulating DNA. In order for characterization to take place a method to clone
total circulating DNA was developed and standardized and thirty-five clones were
obtained and analyzed. It was found that the sequences contain a large amount
of Alu repeats and the significance of this has not been determined yet. This is a
first step towards future studies. / Thesis (M.Sc. (Biochemistry))--North-West University, Potchefstroom Campus, 2007
| Identifer | oai:union.ndltd.org:NWUBOLOKA1/oai:dspace.nwu.ac.za:10394/1329 |
| Date | January 2006 |
| Creators | Van der Vaart, Maniesh |
| Publisher | North-West University |
| Source Sets | North-West University |
| Detected Language | English |
| Type | Thesis |
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