Biology

Study of transfer and persistence of DNA from semen-stained clothing to unstained clothing,in a laundry basket prior to laundering

Having data to give estimates on the properties of DNA as physical evidence will enable investigators and scientists to assess its value as evidence in a particular case. Combining the particular case circumstances with experimental data on these properties will provide a more grounded, Bayesian-type interpretation of a trace DNA profile.

Assessing whether or not DNA can be passively transferred from semen stained garments to unstained garments that are in direct contact prior to being laundered.

Seeking a partner to examine the likelihood of passive DNA transfer between semen-stained and unstained clothing in direct contact in a laundry basket. The variables of interest in this study are (a) time in contact, (b) contact pressure, (c) fabric type (i.e. high absorbency vs. low absorbency) and (d) wet vs. dry fabrics. Working with the CFS project lead, you will develop the experimental design, conduct the experiment and publish the results.Transfer will be measured with bodily fluid testing (AP, P30, microscope) and DNA analysis. If transfer is found to occur the extent of transfer will be assessed.

Microscope, along with standard DNA extraction, quantitation, separation and analysis instruments. The partner must have prior experience with bodily fluid testing and DNA analysis.

CFS will provide reagents for bodily fluid testing and DNA extraction, quantitation and analysis kits. Instrumentation may be provided if needed, or research may be performed in the CFS R&D suite.

Develop a method using InnoTyper 21 to obtain forensically relevant results from degraded, inhibited and low-copy number samples such as bones and rootless hair shaft samples.

Recently a new nuclear DNA typing system, InnoTyper 21 has become available that has been designed to obtain informative results from very degraded, inhibited and low-level forensic samples, including bones and rootless hair shafts. The CFS is currently unable to perform any DNA testing on rootless hair shafts and this kit provides a discriminating alternative to mitochondrial testing.

The goal of this research proposal is to determine an optimized nuDNA extraction method for rootless hair shafts that may permit successful DNA analysis using the InnoQuant HY and InnoTyper 21 kits.

We are seeking an external partner to evaluate whether or not 25 to 50pg of total genomic DNA can be successfully extracted from rootless hair shafts and quantified using the InnoQuant HY kit.

The preferred methods for extraction would be via Qiagen EZ1 (preferred) or organic protocol using phenol/chloroform. Working with the CFS project lead the external partner will develop the experimental design, execute and publish the results. The external partner must have expertise with DNA analysis including extraction, amplification and electrophoresis and will provide instrumentation including (Qiagen EZ1 Advanced XL, Bio-Rad CFX96 or AB 7500 SDS, and AB 3130 or 3500 Capillary Electrophoresis) and needed specialised lab equipment.

The CFS will provide consumables and all required kits including Qiagen-EZ1, InnoQuant and InnoTyper 21 kits.

Study of transfer and persistence of DNA from a person's mouth to other individuals and objects

Having data to give estimates on the properties of DNA as physical evidence will enable investigators and scientists to assess its value as evidence in a particular case. Combining the particular case circumstances with experimental data on these properties will provide a more grounded, Bayesian-type interpretation of a trace DNA profile.

Assessing the extent to which DNA can be passively transferred from items that come in contact with an individual’s mouth (e.g. drinking glasses, cigarettes) to other individuals and objects. The study will focus on primary, secondary and tertiary transfers of DNA.

Seeking a partner to examine the likelihood of passive DNA transfer in the following scenarios. One individual (person 1) will either drink from a glass/cup/bottle or will smoke a cigarette (item 1) and then pass or discard the item, which will then be handled by a second individual (person 2) for a short period of time. The second individual will subsequently handle a “clean” item (item 2) that has not been handled by anyone, or will touch the skin of a third person.

• Primary DNA transfer: Person 1 to item 1
• Secondary DNA transfer: Person 1 to Person 2 (via item 1)
• Tertiary DNA transfer: Person 1 to Item 2 (via Item 1 and Person 2)
• Tertiary DNA transfer: Person 1 to Person 3 (via Item 1 and Person 2)

• Person 2 to Item 2
• Person 2 to Person 3

A sufficient number of individuals and scenarios should perform these activities to obtain robust data that determines the extent and variability of primary, secondary and tertiary DNA transfer in each scenario.

The partner must have prior experience with bodily fluid testing and DNA analysis and have access to the DNA extraction (DNA IQ or DNA Investigator’s kit), quantitation (able to run Plexor HY), separation (capillary electrophoresis) and analysis (compatible with GeneMapper IDX) instruments utilized by the CFS Biology Section. The research partner is expected to design the experimental protocol (in partnership with the CFS project lead), conduct the research project and be lead author on a publication.

The CFS will provide DNA extraction, quantification and separations kits if required. The CFS project lead will contribute to the experimental design and authorship of the expected publication.

Gain a better understanding of DNA trace evidence transfer and persistence patterns using real-time PCR/qPCR to determine quantity and quality of DNA after potential transfer events and stability over time. Specifically, to measure the extent of transfer events (how often trace evidence is detected to be transferred and in what situations transfer occurs) between two fabrics and its detection over time (persistence). The method would first be validated and optimized using a known solution of purified DNA and then additional more complex sources (blood, buccal swab/saliva… etc.) would be analyzed.

Study Transfer and Persistence of DNA from wet, dry or rehydrated bodily fluid stains on porous and non-porous surfaces.

Having data to give estimates on the properties of DNA as physical evidence will enable investigators and scientists to assess its value as evidence in a particular case. Combining the particular case circumstances with experimental data on these properties will provide a more grounded, Bayesian-type interpretation of a trace DNA profile.

Assessing the extent to which DNA from wet, dry or rehydrated bodily fluids stains can be transferred from one substrate to another through contact.

Seeking a partner to examine the likelihood of DNA transfer between bodily fluid-stained and unstained substrates. Blood, semen and saliva stains would be deposited on a variety of porous and non-porous fabrics and surfaces. In various combinations, these same clean fabric and surface types (dry) would be pressed in contact with the wet stain. A second experiment would be carried out similarly with stains being dry and a third experiment performed with dried stains where the contacting fabric/surface is dampened. The amount of time and pressure of contact and the degree of dampness of the contacting surface/fabric would be kept consistent. The source and contacting fabric/surface would be sampled (c/o or swab as applicable) for DNA. Data would be analyzed to compare the starting and transferred amounts of DNA. Working with the CFS project lead, you will develop the experimental design, conduct the experiment and publish the results.

The external partner must have expertise with DNA analysis including extraction, amplification and electrophoresis. Standard DNA extraction, quantitation, separation and analysis instrumentation.

DNA extraction, quant and separations kits. Instrumentation may be provided if needed, or research may be performed in the CFS R&D suite. Training in DNA analysis techniques can be provided by CFS staff.

Assess the value of DNA evidence in Property Crimes during both the police investigations and the during the court litigation process.

The CFS currently measures the value of its services to clients in terms of time to complete reports or the number of reports delivered to clients relative to previously set targets. The CFS is seeking to develop tools to measure the impact of it’s services on the investigative process and on the subsequent litigative process.

Identify one or more measures that demonstrate whether or not forensic DNA analysis has an impact on the police investigation of property crime. For example, does DNA analysis in property crime casework increase or decrease the police resources required to undertake the investigation, or does it shorten or lengthen the investigation. Identify one or more measures that demonstrate whether or not forensic DNA analysis has an impact o the litigation process for property crimes. For example, is the trial longer or shorter when DNA evidence is presented? Does DNA evidence increase or decrease the likelihood of a plea prior to trial?

Seeking a partner to work with the CFS project lead to develop the experimental design, conduct the experiment and publish the results. A high-level description of the project follows. Data-mining casework in the High Volume Service: Determining which types of items are most often submitted, and the outcomes of those examinations (e.g. single source or mixed DNA, is a profile uploaded to the National DNA Databank, is a hit recorded to a Convicted Offender and/or to a crime scene, is a comparison sample submitted from a suspect). Data-mining public records and legal documents relating to these offenses: Determining the legal outcome of the DNA results above (Did the DNA evidence assist in identifying a suspect? Was there other evidence? Was there a plea, or verdict in the case? What role did the DNA evidence have? Were there any significant rulings or decisions made relating to the DNA evidence?) Review of transcripts and/or develop interviews/surveys directed at the end user (i.e. investigators, crown attorney)

Upon completion of the data-mining, recommendations regarding best practices can be made – were there patterns observed to suggest certain types of evidence had a bigger/lesser impact than others? The outcome of this research will be a series of recommendations for the optimization of the High Volume Service (used for the analysis of property crime at the CFS)

Expertise in designing and conducting and evaluating longitudinal or associational research.

Case-specific information stored in relational databases.

Study transfer and persistence of DNA deposited onto face, neck and chest surfaces while people are talking in close proximity.

Having data to give estimates on the properties of DNA as physical evidence will enable investigators and scientists to assess its value as evidence in a particular case. Combining the particular case circumstances with experimental data on these properties will provide a more grounded, Bayesian-type interpretation of a trace DNA profile.

To determine whether or not measurable amounts of DNA can be deposited during an intimate (close proximity) conversation onto the face, neck and chest of the partner in the conversation.

Seeking a partner to work with the CFS project lead to develop the experimental design, conduct the experiment and publish the results. A high level description of the project follows. Face (both cheeks) and neck swabs would be taken from all subjects, and unexposed breast swabs and exposed chest swabs would be taken from female subjects only, to assess level of background DNA. The skin would then be cleaned. Male and female pairs would spend varying fixed amounts of time speaking together in an environment replicating a social scenario where voices would be raised and individuals would be in close contact. Face and neck swabs (all subjects) and chest and breast swabs (female subjects) would be taken again following the timed interaction. DNA analysis would be performed on all swabs and results interpreted.

The external partner must have expertise with DNA analysis including extraction, amplification and electrophoresis. Standard DNA extraction, quantitation, separation and analysis instrumentation.

DNA extraction, quant and separations kits. Instrumentation may be provided if needed, or research may be performed in the CFS R&D suite. Training in DNA analysis techniques can be provided by CFS staff.

Can DNA deposited on fabric, as a result of a touch or more vigorous action (e.g. grabbing), be localized and differentiated from DNA deposited through normal wear of the clothing item?

Having data to give estimates on the properties of DNA as physical evidence will enable investigators and scientists to assess its value as evidence in a particular case. Combining the particular case circumstances with experimental data on these properties will provide a more grounded, Bayesian-type interpretation of a trace DNA profile.

To determine whether or not a chemical test can be developed to permit the visualisation of DNA deposited by a touch or by a more vigorous action (e.g. grabbing) To determine whether a chemical test can be developed to permit the visualisation of DNA deposited by a touch or more vigorous action (e.g. grabbing) versus the background DNA from normal wearing of the item.

Working with the CFS project lead, you will develop the experimental design, conduct the experiment and publish the results. We are seeking to develop a chemical testing method for visually localizing touch DNA that (1) is both practical and safe to use in the lab, (2) reliably detects and distinguishes a touch DNA deposition from background sources and (3) allows for the best recovery of DNA.

The external partner must have expertise with DNA analysis including extraction, amplification and electrophoresis. Standard DNA extraction, quantitation, separation and analysis instrumentation.

Alternative light source if required for luminescent visualisation (Projectina Scene of Crime Lamp). DNA extraction, quant and separations kits. Instrumentation may be provided if needed, or research may be performed in the CFS R&D suite. Training in serology and DNA analysis techniques can be provided by CFS staff.

Study transfer and persistence of DNA deposited on the body during sexual activities, which are followed by bathing or showering

Having data to give estimates on the properties of DNA as physical evidence will enable investigators and scientists to assess its value as evidence in a particular case. Combining the particular case circumstances with experimental data on these properties will provide a more grounded, Bayesian-type interpretation of a trace DNA profile.

To study whether or not foreign DNA deposited on skin in various body locations during sexual activities can be detected after showering or bathing.

Seeking a partner to work with the CFS project lead to develop the experimental design, conduct the experiment and publish the results. A high-level description of the project follows. Completion will require consenting individuals to perform sexual activities, followed by collection of bodily samples using swabs. Areas will be swabbed immediately after the sexual activity for the purpose of determining a baseline amount of deposition. Participants should resume their normal activities after the sexual contact has occurred. They should record how much time has elapsed between the sexual contact and the shower/bath. Following collection of the swabs, DNA analysis including extraction, quantification (total and male DNA), amplification using Identifiler plus and interpretation following CFS protocols is required. A sufficient number of individuals should perform these activities in order to obtain robust data that determines the extent of persistence of foreign DNA in each scenario.

The external partner must have expertise with DNA analysis including extraction, amplification and electrophoresis. Standard DNA extraction, quantitation, separation and analysis instrumentation.

DNA extraction, quant and separations kits. Instrumentation may be provided if needed, or research may be performed in the CFS R&D suite. Training in DNA analysis techniques can be provided by CFS staff.

Study the transfer and persistence of DNA from habitually worn clothing relative to the most recent wearer of an item.

Having data to give estimates on the properties of DNA as physical evidence will enable investigators and scientists to assess its value as evidence in a particular case. Combining the particular case circumstances with experimental data on these properties will provide a more grounded, Bayesian-type interpretation of a trace DNA profile.

To assess whether or not the DNA results derived from different sampling methods would permit conclusions to be drawn about the habitual wearer of an item vs the most recent wearer of an item.

Seeking a partner to examine three classes of items: Those worn and rarely washed; those worn but not washed after every wear; those worn and washed after one wear. All items should be purchased new and laundered prior to use in the study. Note that working with the CFS project lead, you will develop the experimental design, conduct the experiment and publish the results.

The following is a high-level description of the experimental design.

1. Items that are worn and not regularly washed:

Individual 1 wears for 30 days and Individual 2 then wears for one day. Sampling is as described: Toques – cut out from front inside headband followed by swab of entire inside headband. Baseball hats – cut out from front inside headband followed by swab of entire inside headband. Fabric gloves – cut out from thumb pad followed by swab of inside surface of all fingers.

2. Items that are worn and not washed after every wear:

Individual 1 wears for 5 days; Individual 2 then wears for one day. Sampling as described. Pants/jeans - Sample waistband by fabric cutting inside back waist, then swab entire waist. Also could take a fabric cutting from front pocket at pocket opening and then a swab of the entire pocket opening

3. Items that are worn and washed regularly, typically after one wear:

Individual 1 wears for 1 day; individual 2 then follows up by wearing for 1 day. Sampling as described. T-shirts/tank tops - Sample back inside neckline by first a fabric cutting from the back and then swabbing the entire neckline.

The DNA profiles determined would be compared to known comparison samples of the participants to determine total amounts of DNA and which wearer was the major contributor vs the minor contributor.

The external partner must have expertise with DNA analysis including extraction, amplification and electrophoresis. Standard DNA extraction, quantitation, separation and analysis instrumentation.

DNA extraction, quant and separations kits. Instrumentation may be provided if needed, or research may be performed in the CFS R&D suite. Training in DNA analysis techniques can be provided.

1. Use single source samples to develop criteria for identification of body fluids and body sources of epithelium.
2. Determine whether the cell composition of mixtures determined by IFC align with the mixture proportions obtained with DNA analysis.
3. Characterize single source and mixtures of body fluids/epithelium on commonly encountered forensic case work items.
4. Determine whether IFC accurately identifies the time since deposition of body fluids and body source epithelium.

Sexual assault samples contain DNA from multiple sources, making them one of the most challenging types of forensic samples to analyze, often beyond the reach of rapid DNA analyzers. The goal of this project is to develop a workflow that facilitates the use of rapid DNA typing instruments for all samples, including those collected in sexual assaults.

(1) Enable processing of most sexual assault samples automatically (2) Enable the generation full STR profiles from degraded samples

University of Toronto researchers have indicated interest in the development of a microfluidic platform for the separation of different cell types in forensic samples commonly encountered in sexual assault investigations

Access to CFS forensic processing capabilities, including, equipment (EZ1 Advanced XL automated liquid handler, thermomixer, qPCR), software (Genemapper IDX, Probabilistic STARmix), staff resources and consumables as required.