The Effect of Sample and Sample Matrix on DNA Processing: Mechanisms for the Detection and Management of Inhibition in Forensic Samples

The presence of inhibitory substances in biological forensic samples has, and continues to affect the quality of the data generated following DNA typing processes. Although the chemistries used during the procedures have been enhanced to mitigate the effects of these deleterious compounds, some challenges remain. Inhibitors can be components of the samples, the substrate where samples were deposited or chemical(s) associated to the DNA purification step. Therefore, a thorough understanding of the extraction processes and their ability to handle the various types of inhibitory substances can help define the best analytical processing for any given sample. A series of experiments were conducted to establish the inhibition tolerance of quantification and amplification kits using common inhibitory substances in order to determine if current laboratory practices are optimal for identifying potential problems associated with inhibition. DART mass spectrometry was used to determine the amount of inhibitor carryover after sample purification, its correlation to the initial inhibitor input in the sample and the overall effect in the results. Finally, a novel alternative at gathering investigative leads from samples that would otherwise be ineffective for DNA typing due to the large amounts of inhibitory substances and/or environmental degradation was tested. This included generating data associated with microbial peak signatures to identify locations of clandestine human graves. Results demonstrate that the current methods for assessing inhibition are not necessarily accurate, as samples that appear inhibited in the quantification process can yield full DNA profiles, while those that do not indicate inhibition may suffer from lowered amplification efficiency or PCR artifacts. The extraction methods tested were able to remove >90% of the inhibitors from all samples with the exception of phenol, which was present in variable amounts whenever the organic extraction approach was utilized. Although the results attained suggested that most inhibitors produce minimal effect on downstream applications, analysts should practice caution when selecting the best extraction method for particular samples, as casework DNA samples are often present in small quantities and can contain an overwhelming amount of inhibitory substances.

The effect of sample and sample matrix on DNA processing: Mechanisms for the detection and management of inhibition in forensic samples

The presence of inhibitory substances in biological forensic samples has, and continues to affect the quality of the data generated following DNA typing processes. Although the chemistries used during the procedures have been enhanced to mitigate the effects of these deleterious compounds, some challenges remain. Inhibitors can be components of the samples, the substrate where samples were deposited or chemical(s) associated to the DNA purification step. Therefore, a thorough understanding of the extraction processes and their ability to handle the various types of inhibitory substances can help define the best analytical processing for any given sample. A series of experiments were conducted to establish the inhibition tolerance of quantification and amplification kits using common inhibitory substances in order to determine if current laboratory practices are optimal for identifying potential problems associated with inhibition. DART mass spectrometry was used to determine the amount of inhibitor carryover after sample purification, its correlation to the initial inhibitor input in the sample and the overall effect in the results. Finally, a novel alternative at gathering investigative leads from samples that would otherwise be ineffective for DNA typing due to the large amounts of inhibitory substances and/or environmental degradation was tested. This included generating data associated with microbial peak signatures to identify locations of clandestine human graves. Results demonstrate that the current methods for assessing inhibition are not necessarily accurate, as samples that appear inhibited in the quantification process can yield full DNA profiles, while those that do not indicate inhibition may suffer from lowered amplification efficiency or PCR artifacts. The extraction methods tested were able to remove >90% of the inhibitors from all samples with the exception of phenol, which was present in variable amounts whenever the organic extraction approach was utilized. Although the results attained suggested that most inhibitors produce minimal effect on downstream applications, analysts should practice caution when selecting the best extraction method for particular samples, as casework DNA samples are often present in small quantities and can contain an overwhelming amount of inhibitory substances.^

Reading comprehension and mathematical concept acquisition through the use of math stories with bilingual children

Math storybooks are picture books in which the understanding of mathematical concepts is central to the comprehension of the story. Math stories have provided useful opportunities for children to expand their skills in the language arts area and to talk about mathematical factors that are related to their real lives. The purpose of this study was to examine bilingual children's reading and math comprehension of the math storybooks. ^ The participants were randomly selected from two Korean schools and two public elementary schools in Miami, Florida. The sample consisted of 63 Hispanic American and 43 Korean American children from ages five to seven. A 2 x 3 x (2) mixed-model design with two between- and one within-subjects variable was used to conduct this study. The two between-subjects variables were ethnicity and age, and the within-subjects variable was the subject area of comprehension. Subjects were read the three math stories individually, and then they were asked questions related to reading and math comprehension. ^ The overall ANOVA using multivariate tests was conducted to evaluate the factor of subject area for age and ethnicity. As follow-up tests for a significant main effect and a significant interaction effect, pairwise comparisons and simple main effect tests were conducted, respectively. ^ The results showed that there were significant ethnicity and age differences in total comprehension scores. There were also age differences in reading and math comprehension, but no significant differences were found in reading and math by ethnicity. Korean American children had higher scores in total comprehension than those of Hispanic American children, and they showed greater changes in their comprehension skills at the younger ages, from five to six, whereas Hispanic American children showed greater changes at the older ages, from six to seven. Children at ages five and six showed higher scores in reading than in math, but no significant differences between math and reading comprehension scores were found at age seven. ^ Through schooling with integrated instruction, young bilingual children can move into higher levels of abstraction and concepts. This study highlighted bilingual children's general nature of thinking and showed how they developed reading and mathematics comprehension in an integrated process. ^