Preliminary Study: DNA Transfer and Persistence on Non-Porous Surfaces Submerged in Spring Water.

Submerged items are often thought to lack evidentiary value. However, previous studies have shown the ability to recover DNA from submerged porous items for upwards of six weeks. The crevices or interweaving fibers in porous items are thought to protect DNA from being washed away. It is hypothesized that, because non-porous surfaces do not have the same traits that might aid in DNA retention, then DNA quantities and the number of donor alleles recovered would decrease over longer submersion periods. Additionally, it is hypothesized that DNA quantity and the number of alleles would be negatively affected by flow conditions. Neat saliva of known DNA quantity was applied to glass slides and exposed to stagnant and flowing spring water to observe the effects on both DNA quantity and STR detection. Results supported that DNA deposited onto glass and subsequently submerged in water experienced a decrease in DNA quantity over time, yet submersion did not have as strong of a negative effect on the detected amplification product. Additionally, an increase in DNA quantity and detected amplification product from designated blank slides (no initial DNA added) could indicate the possibility of DNA transfer.

Assessment of PowerPlex® Fusion 5C's ability to type degraded DNA.

DNA samples collected at crime scenes are often degraded so this research focused on the ability of the Promega PowerPlex® Fusion 5C amplification kit to type both naturally and artificially degraded DNA. DNA was degraded naturally by placing equal volumes of blood on white fabric that was stored either inside, outside in a shaded area, or outside in direct sunlight. Samples were then collected every 10 days for 60 days and the DNA extracted (QIAamp® DNA Investigator). Artificially degraded samples were created by exposing extracted DNA to either UV light or 95 °C heat for varying times. DNA was also degraded artificially by placing blood samples into a 50% bleach solution for varying times prior to extraction. Following sample treatment, standard forensic DNA analysis was performed including quantification (Investigator® Quantiplex) and amplification (PowerPlex® Fusion 5C). Separation and detection were performed on an ABI 3130xl capillary electrophoresis unit and analysis was performed using GeneMapper ID v3.2.1. While the time and shade samples showed similar amounts of degradation, the samples exposed to direct sun showed more degradation. The artificially degraded samples showed more signs of degradation such as reduced overall peak height and peak height imbalance at heterozygous loci. There were also some cases where an allele that was known to be in the profile exhibited total dropout. Although there were some instances of both allelic dropout and heterozygote peak imbalance, PowerPlex® Fusion was able to reliably type degraded DNA as all alleles detected were consistent with the known donor profile. The results show that PowerPlex® Fusion is a robust kit capable of handling forensically challenged samples.

A preliminary assessment of the effect of PreCR™ DNA repair treatment on mixture ratios in two person mixtures.

In this study, DNA extracted from known buccal samples was combined into two component mixture samples. These were subjected to UV exposure prior to their amplification with the Promega PowerPlex® 16HS amplification kit, and subsequent capillary electrophoresis on the ABI 3130xl instrument. Damaged samples were subjected to enzymatic repair treatment and retested to assess the amount of repair. Data showed that there is fidelity associated with the application with profile concordance after its use, and a corresponding increase in the amount of recovered alleles post damage. Results also showed changes in the stochastic relationship between mixture components that appear to be induced by the repair process itself. The mixture ratios of DNA samples were altered from an approximate original 1:3 ratio, to a ratio of 1:2 or greater. This variation can have a significant effect regarding the ability to reliably de-convolute DNA mixtures that have been subjected to the repair process.