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| Auteur||K.D. van den Bogaert|
|Titel||EFFECTS OF BSA AND COLLAGENASE ON DNA PROFILING FROM BONE|
|Begeleiders||Dennis McNevin, Ate Kloosterman|
|Faculteit||Faculteit der Natuurwetenschappen, Wiskunde en Informatica|
|Opleiding||FNWI MSc Forensic science|
|Samenvatting||DNA-typing technologies are instrumental in the identification of victims from a mass disaster. While DNA profiling can provide a very high probability of identity, it is not always successful for disaster victim identification. Often only bone and teeth remain and these tissues pose the problem of low quantities of DNA and the possible presence of PCR inhibitors. These can make DNA profiling and thus identification of the remains impossible. To overcome this problem the effect of two additions to the standard Austrian Federal Police (AFP) method for DNA profiling from bone were investigated by tests with bovine bone samples.|
The addition of BSA is often mentioned as an addition to the PCR that will increase DNA yield. However, the underlying mechanism is unknown. This research tested whether the positive effect of BSA depends on the presence of inhibitors. It was found that of the tested alleged inhibitors (EDTA, SDS, collagen, humic acid, and tannic acid) only humic acid, EDTA and SDS inhibited the PCR and that BSA only has a positive effect on the DNA yield when humic acid is present. Thus the positive effect depends on the presence of this inhibitor.
The addition of collagenase to the extraction buffer is thought to lead to the breakdown of the alleged inhibitor collagen. This should have the dual advantage of breaking down the bone’s physical barrier that protects its DNA, and the (partial) elimination of the inhibitor. It can be concluded that collagenase has a slightly positive effect on the DNA yield. However, this is only the case in clean DNA samples and not in bone samples. However, it has been demonstrated that collagenase is able to reverse the inhibitory effects of collagen when the collagenase protocol was used.
When using actual bone samples the performance of the individual BSA or collagenase protocols did not lead to a significantly higher DNA yield compared to the standard AFP protocol. The combined protocols however do lead to a significant increase of the DNA yield when compared to this standard.
|Soort document|| scriptie master|
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