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Validation of the M-Vac® cell collection system and examination of DNA extraction methods using the EZ1 biorobotGlater, Moss Samuel 01 February 2023 (has links)
The ability to generate deoxyribonucleic acid (DNA) profiles from genetic material is a powerful tool commonly used by forensic scientists. The first major step of this process is the collection of DNA from a crime scene. DNA may be deposited in many ways, including in biological fluids and by touch DNA deposition. Techniques such as swabbing are commonly used, but often pose problems when used on rough, porous surfaces. The M-Vac® is a wet-vacuum designed to spray and recollect a buffer solution from a surface to allow for the collection of DNA from these porous objects.
Dilute saliva samples and touch DNA via handling were deposited on assorted items and collected using both traditional swabbing and wet vacuum methods. Further, M-Vac® filters containing collected material were extracted using the EZ1 Biorobot either in a single tube or divided into approximately quarters and extracted in four separate tubes to assess DNA yields from both methods. The M-Vac® was shown to collect equal or greater amounts of DNA compared to swabbing from various porous surfaces, and the samples collected with the M-Vac® were shown to produce more complete DNA profiles. Use of the M-Vac® on surfaces following swab collection resulted in the recovery of additional DNA, often exceeding the amount initially recovered from the swab. It was also observed that in nearly all profiles generated from touch DNA sources, alleles were detected that originated from a non-donor source. In some cases, extraction from filters in quarters resulted in a higher total amount of DNA than whole filters when all four quarters were added together. However, quarters from a single filter had a high amount of variation in the level of DNA, indicating that the DNA may not be distributed evenly on the filter.
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Evaluation of a genomic work flow for the detection of Bacillus subtilis in animal feed and food samplesLindberg, Stina January 2005 (has links)
<p>Bacillus anthracis is one of the most feared agents of biological warfare and causes the</p><p>deadly disease called anthrax. SVA (statens veterinärmedicinska anstalt) is working on a</p><p>project together with SLV (statens livsmedelsverk) where the target is to find rapid and</p><p>effective detection methods for Bacillus anthracis in animal feed and food samples. Bacillus</p><p>subtilis, which is harmless, was used in this study as a model organism to Bacillus anthracis.</p><p>A known concentration of vegetative Bacillus subtilis was spiked in animal feed and food</p><p>samples. The genomic work flow was based on automated DNA isolation and real time PCR.</p><p>The aim of the study was to screen for inhibitory components in the animal feed and food</p><p>samples using two different DNA isolation robots; Magnatrix 8000 and Biorobot EZ1. The</p><p>results showed that DNA of high quality was extracted from the samples with both robots.</p><p>However, the CT-value generated by the real time PCR showed considerable variation</p><p>depending on the sample matrix. Some samples, for instance egg and liver, were problematic</p><p>and gave low concentrations and high CT-values probably due to inhibitory components in the</p><p>samples. Further studies will be needed to solve these problems and optimize the methods that</p><p>were used in this study.</p>
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Evaluation of a genomic work flow for the detection of Bacillus subtilis in animal feed and food samplesLindberg, Stina January 2005 (has links)
Bacillus anthracis is one of the most feared agents of biological warfare and causes the deadly disease called anthrax. SVA (statens veterinärmedicinska anstalt) is working on a project together with SLV (statens livsmedelsverk) where the target is to find rapid and effective detection methods for Bacillus anthracis in animal feed and food samples. Bacillus subtilis, which is harmless, was used in this study as a model organism to Bacillus anthracis. A known concentration of vegetative Bacillus subtilis was spiked in animal feed and food samples. The genomic work flow was based on automated DNA isolation and real time PCR. The aim of the study was to screen for inhibitory components in the animal feed and food samples using two different DNA isolation robots; Magnatrix 8000 and Biorobot EZ1. The results showed that DNA of high quality was extracted from the samples with both robots. However, the CT-value generated by the real time PCR showed considerable variation depending on the sample matrix. Some samples, for instance egg and liver, were problematic and gave low concentrations and high CT-values probably due to inhibitory components in the samples. Further studies will be needed to solve these problems and optimize the methods that were used in this study.
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