Social and Behavioral Sciences Commons^™

Development And Validation Of Microrna Markers For Forensic Body Fluid Identification Using Rt-Qpcr, Jane Goble

Master of Science in Forensic Science Directed Research Projects

Body fluid identification is one of the first and most fundamental steps in the forensic DNA workflow. It allows analysts to determine where a sample might exist on an item of evidence, as well as the type of sample that they are dealing with. However, current serological methods have been documented to produce false or inaccurate results in some cases, and may consume large amounts of sample during processing. Molecular methods, such as the use of microRNA and microbial profiles have been introduced as a means to increase specificity of testing, and have even been combined into a singular assay …

Development Of A Loop-Mediated Isothermal Amplification (Lamp) Assay For Detection Of Mirnas, Alisha B. Whelan

Master of Science in Forensic Science Directed Research Projects

Body fluid identification serves a vital role in contextualizing biological evidence received in forensic laboratories. While several fluids can be presumptively detected using immunological or catalytic tests, these tests are susceptible to false results and have limited sensitivity. True confirmatory testing is still somewhat limited in forensic laboratories. With the advancement of DNA typing technologies, laboratories can develop DNA profiles from trace amounts of sample, increasing the demand for sensitive and accurate body fluid detection. This project proposes using loop-mediated isothermal amplification (LAMP) as a method to rapidly detect microRNAs (miRNAs) that are differentially expressed in various body fluids with …

Stability And Variation Within Donors Of Mirna Markers For Body Fluid Identification, Kelsey R. Price

Master of Science in Forensic Science Directed Research Projects

Traditional enzymatic tests for body fluid identification are prone to false negatives, false positives, and several body fluids do not have confirmatory or reliable presumptive tests. Therefore, molecular-based tests may be more reliable, such as messenger RNA (mRNA), microbial DNA, or microRNA (miRNA) assays.

miRNAs are small, noncoding RNAs whose main function in-vivo is the regulation of protein expression by selectively suppressing the translation of their corresponding mRNAs. They also lack a poly-A tail, and because of this, their small size, and their association with other molecules such as the RISC protein complex, they are more robust than other RNAs, …

Impact Of 1,4-Dithiothreitol (Dtt) On Microbial Community Structures Associated With Human Semen Samples, Christian R. Blaise

Master of Science in Forensic Science Directed Research Projects

Body fluid identification (BFID) is one of the most important steps in forensic science as it provides important context to how DNA analysis should be performed on biological samples found at the scene of a crime. Many scientific methods on BFID have been developed over the years with respective benefits and flaws. One emerging method in recent years is microbial signature based BFID. This method uses the same DNA extract that is utilized for human DNA profile generation for microbial analysis. Although DNA is extracted from all body fluid samples without any additional steps, extraction of human DNA from semen …

Effects Of Microwave Radiation On The Digestion Of Proteins Involved In Body Fluid Identification, Aaron J. Fox

Student Theses

Body fluid identification plays an important role in understanding how the events during a crime may have taken place. The presence of a body fluid may help identify an individual who committed a crime, while the type of body fluid present may help investigators determine how a crime occurred. Current body fluid identification techniques are not always conclusive and may only suggest the presence of a body fluid type. A protocol developed by the New York City Office of the Chief Medical Examiner (OCME) established a method that confirms body fluid type through mass spectrometry. To identify marker proteins in …

Semi-Quantitative Detection Of Signature Peptides In Body Fluids By Liquid Chromatography Tandem Mass Spectrometry (Lc-Ms/Ms), Tebah N. Browne

Student Theses

This study covers a modified semi-quantitative approach for liquid chromatography mass spectrometry (LC-MS/MS) signature peptide detection for body fluid identification. Peptide concentrations were measured based on synthetic peptide standards. Samples were processed with varying trypsin digestion and purification protocols, including a three-hour trypsin digestion and Microcon membrane filtration. The Microcon filtration method generates polymerase chain reaction (PCR) compatible DNA and peptide fractions that can be analyzed without any further purification. Preliminary validation tests covered stains on different substrates, semen/ saliva mixtures, minimum sample volume, and repeatability. All signature peptides in the multiplex were present at different concentrations and varied amongst …

Investigating The Isolation And Amplification Of Micrornas For Forensic Body Fluid Identification, Kelsie R. O'Leary, Claire L. Glynn

Forensic Science Publications

Background: The discovery of forensic DNA typing evolved molecular biology far beyond what could have been expected in terms of its forensic application, and now there exists other developments in molecular biology which are ready for application to forensic challenges. One such challenge is the identification of the body fluid source of stains recovered from evidence items and crime scenes. Currently there are significant efforts in the research field to develop novel methods for the molecular identification of body fluids, with microRNAs (miRNAs) revealing great potential. MiRNAs have been shown to have high tissue specificity and are less susceptible to …