Abstract
Circulating DNA is comprised of fragments of genomic and mitochondrial DNA that can be isolated from the blood and body fluids of vertebrates. The potential use of these fragments as biomarker(s) for cancer and other diseases has been investigated extensively, but still the exact mechanism of origin and the function or significance of these molecules have not been explained. We attempted to elucidate this question by means of parallel tagged sequencing of circulating DNA on the GSFLX sequencer from 454 life sciences. Circulating DNA from 22 consenting individuals of which 12 were prostate cancer patients and 10 were controls were sequenced simultaneously on one 16th of a picotiter plate. A total of ~3,600 unique sequences were analyzed, these were distributed over the human genome and a slightly higher mutation rate could be observed for DNA obtained from the cancer patients compared to the control group. We concluded that the distribution of circulating DNA over the genome is similar for both groups even though some sequence related differences might be observed. Circulating DNA can be seen as a representation of genomic DNA in the blood of an individual.
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References
Lander ES, Linton LM, Birren B et al (2001) Initial sequencing and analysis of the human genome. Nature 409:860–921
Meyer M, Stenzel U, Myles S et al (2007) Targeted high-throughput sequencing of tagged nucleic acid samples. Nucleic Acids Res 35:e97
Scherer S (2008) A short guide to the human genome, 1st edn. Cold Spring Harbor Laboratory Press, New York, NY
van der Vaart M, Pretorius PJ (2010) Is the role of circulating DNA as a biomarker of cancer being prematurely overrated? Clin Biochem 43:26–36
van der Vaart M, Semenov DV, Kuligina EV et al (2009) Characterization of circulating DNA by parallel tagged sequencing on the 454 platform. Clin Chim Acta 409:21–27
Acknowledgments
This work was partially supported by The National Research Foundation of South Africa (MvdV), The Russian Federal Agency for Science and Innovations 02.522.12.2005 (DVS), RFBR 10-04-01386-a, 10-04-01442-a (DVS) and by SB RAS Grant 18 (DVS).
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van der Vaart, M., Semenov, D.V., Kuligina, E.V., Richter, V.A., Pretorius, P.J. (2010). Parallel Tagged 454 Sequencing for the Characterization of Circulating DNA. In: Gahan, P. (eds) Circulating Nucleic Acids in Plasma and Serum. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9382-0_4
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DOI: https://doi.org/10.1007/978-90-481-9382-0_4
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