99th SEMI-ANNUAL SEMINAR (Spring 2002)
May 7-11, 2002
San Francisco, California

Peter D. Barnett Forensic Science Associates

A multidisciplinary investigation can be compared with a good pizza dinner. A good pizza requires quality raw ingredients, proper equipment, and skilled cooks. But with only those ingredients the dinner could easily end up being a plateful of red-sauce-covered, gooey dough, with too many anchovies, no pepperoni, and the whole mess cold in the center-not a meal that is inclined to make the discriminating diner leave a big tip and come back a second time. What the good pizza requires is a good chef-the person who has an understanding of what the final pizza should look and taste like, can select the raw ingredients, can combine them appropriately to make the dough, the sauce, and the cheese. From the long list of available ingredients, anchovies to zucchini and goat to gorgonzola, the chef must select which ones will go together to make an acceptable final result. The chef may assign certain tasks to subordinates-pot scrubbers, sous chefs, or even ingredient suppliers-but the chef bears the ultimate responsibility for the final product. A not uncommon mistake is to give all of the credit, or the blame, to the food server-which is akin to blaming the UPS driver when your mail order shoes don't fit. A modern, multidisciplinary forensic science investigation is not different except for the fact that the forensic equivalent of the chef is a lawyer who is generally unable to recognize quality ingredients or appropriate combinations, but is more or less adept at convincing the diners(the jurors) that the shapeless, soggy mess on their platter is convincing proof of a defendant's guilt or innocence. Just as a pizza parlor requires the direction of a good chef to produce good pizza, and a forensic science investigation requires the direction of a good scientist to produce a good result. Producing either a good pizza or a good investigation requires a person, the chef or the forensic scientist, who is knowledgeable in all phases of the process. Without that person, the best that one can hope for is a mess that no one will buy. At worst one might expect a pizza or investigation that could make someone dead. Combining reality with allegory, the forensic investigation of a pizza will be presented. This presentation will demonstrate that the forensic investigation of a pizza can be just as complicated, involve just as many types of evidence, and require just a broad a range of skills and abilities as any forensic investigation. The complexities of a forensic investigation of a pizza can involve a variety of disciplines and different types of evidence - more, perhaps, that the ingredients of the pizza itself. The importance of the forensic scientist in conducting the investigation is no less than the importance of the chef in the preparation of the pizza.

Martha Blake San Francisco Police Department

On December 30, 2000, two male juveniles were arrested for grand theft of animals (two koala bears) from the San Francisco Zoo. Fortunately, the marsupials were recovered, scared but safe. One of the two juveniles confessed to the burglary but the other juvenile was not as cooperative. The black wool jacket worn by this juvenile was examined for the presence of foreign hairs. Numerous wavy gray hairs were recovered from this jacket and examined. These hairs were microscopically compared to koala bear hairs submitted by the zookeeper and to other marsupial hairs for comparison. Numerous hairs removed from the jacket were identified morphologically as koala bear hairs. Of particular significance is the scale pattern of these hairs. Koala bear hairs have unique identifying features, and unless you are a zookeeper, it is hard to explain why these hairs are on your clothing.

Lowell W. Bradford

Describes a firearms homicide with an unusual combination of investigative procedure coupled with laboratory coordination involving firearms and document evidence, resulting in a determinative identification of the assailant.

Dawn T. Bravo*, Stanley M. Parsons and David O. Harris. Department of Chemistry and Biochemistry, University of California, Santa Barbara

The objective was to develop an inexpensive, sensitive, rapid and selective color test for GHB (4-hydroxybutyrate). The test should not produce false positive or negative results from a wide range of sample types. Such a test would provide means by which (i) people can protect themselves from unknowingly ingesting GHB, (ii) law enforcement can obtain probable cause to detain persons possessing GHB, (iii) forensic laboratories can screen suspect samples before definitive testing, and (iv) emergency rooms can screen bodily fluid for rapid diagnosis. The enzyme GHB dehydrogenase reacts GHB with the cofactor NAD+ to produce NADH. Dehydrogenase was cloned from Ralstonia eutropha and expressed in Escherichia coli as a readily purified fusion protein. The commercial enzyme diaphorase reacts NADH with colorless tetrazolium prodye to form a strong dye. This reaction coupled to the dehydrogenase reaction produces intense color when GHB is present; however, ethanol is an alternative substrate of the dehydrogenase, and it can produce a false positive with untainted alcoholic drinks. Nevertheless, ethanol binds 200-fold more weakly to the enzyme; this provides a theoretical basis to detect GHB preferentially. Filter paper carrying covalently bound fusion protein and a large excess of diaphorase (so dehydrogenase controls the rate of color formation), NAD+ and tetrazolium prodye rapidly becomes intensely colored in 176 mg GHB/ml (2 mM) but not in aqueous ethanol. False positives can be avoided more economically by evaporation of the suspect sample to remove ethanol before testing. Evaporation of very small volumes of twenty-five common bar drinks and urine on filter paper followed by application of test reagents to the paper detected as little as 8.8 mg GHB/ml (0.1 mM). Inexpensive, sensitive, rapid and selective color tests for GHB in a wide range of sample types have been developed.

Bruce Broce Comisión de la Verdad de Panamá, Rep. de Panamá

A decade after U.S. Armed Forces had dismantled Panama's military apparatus and many Panamanians had decided to move forward instead of looking back, the specter of the dictatorship was summoned by the exhumation of two sets of skeletal remains at a former military barracks in September 1999. A little more than a year later, excavations at the same site yielded two additional sets of remains, placing the period of military rule squarely in the crosshairs of a national debate. In part influenced by these events, President Mireya Moscoso announced in early January 2001 that she would be creating a truth commission that would examine gross human rights abuses committed by the military regime during their 21-year reign. On January 18, 2001, Moscoso's presidential decree brought into being la Comisión de la Verdad de Panamá (CVP), or Truth Commission of Panama.

Dianne Burns California Department of Justice, DNA Laboratory

The future of forensic DNA technology is as secure and untouchable as a Van Gogh in a Swiss bank vault. On a global scale, police, politicians, and prosecutors, maintain a ferocious appetite for the individualizing capabilities of our genetic blueprint. The brilliant accomplishments of the DNA revolution have forced other forensic specialties to consider justifying their value to the criminal justice system or accept playing a smaller game. On both sides of the Atlantic trace evidence practitioners are asking the same question: "What is the value of trace evidence in a DNA world?" That is the question this presentation will explore.

Goldie Chopra PM 17, Bhupinder Nagar Road, Near Railway Crossing No. 22, Patiala (Punjab) - 147001 INDIA

The paper deals with the chemical analysis and classification of toners used in photocopying processes by FTIR. The purpose of the work is to: 1) determine if the spectrum of the toner extracted from a photocopy and measured by FTIR is consistent with the corresponding spectrum of the raw toner powder. 2) determine whether IR spectra obtained for raw and processed toner can be differentiated from spectra obtained from toners used in other models of same brand and also from other machines of different brands. 3) if the above is true, then it is possible to devise a classification scheme for classifying toners used in photocopying machines on the basis of characteristic absorption bands in the IR spectra.

Lloyd Cunningham Forensic Document Examiner

Mass killings in 1980's at a Calaveras county cabin. Suspects Leonard Lake and Charles Ng. This presentation will cover the document trail that suspects Leonard Lake and Charles Ng left behind. Forensic document examinations revealed evidence that tied Lake and Ng to the grisly murders, which include identity theft, burial sites, murder plans and after the fact cartoons that depict knowledge of the murders.

Lloyd Cunningham Forensic Document Examiner

This presentation deals with the subject of how trial attorneys prepare-prepare-prepare to cross-examine an expert witness. A "moot court" session was held at the 2001 American Bar Association's litigation section national meeting, where this speaker, serving as an expert witness, was subjected to vigorous cross-examination by four of the top trial attorneys in the country. As part of the "moot court" program, a paper entitled "Cross Examining an Expert Witness: What to Do, What Not to Do" was provided to all attendees. This paper was designed to attack the credibility of all expert witnesses. Excerpts from this paper will be discussed and they will certainly disclose the agenda that is in place to make your visit to the witness stand a very memorable one.

Yasser Daoudi*, Rhonda K. Roby, Susanne Dietz, Tina McIntosh, Yu-Hui Rogers, Matthew Reardon, Timothy Stockwell, and Mark Adams; Applied Biosystems

The City of New York Office of Chief Medical Examiner has orchestrated a team of scientists using DNA technology to identify the victims of the World Trade Center. Applied Biosystems and Celera have been asked to conduct mitochondrial DNA (mtDNA) sequence analysis from extracted DNA from the family reference specimens and the evidentiary material recovered from this disaster site. Mitochondrial DNA analysis is a valuable tool for human remains identification. Due to its high copy number, mtDNA analysis is especially useful when the DNA is highly degraded or when only small sample sizes are available. Additionally, due to its maternal inheritance, mtDNA analysis is a valuable resource when reference standards or close relatives are not available for comparison. Implementation of and advancements in automation for forensic mtDNA analysis in advanced laboratory robotics, multicapillary electrophoretic systems, and data analysis will be presented in light of the World Trade Center Disaster. Data analysis is the largest challenge for mtDNA testing of this magnitude. Clearly, automation in the laboratory has exceeded the automation for data analysis. Efforts to reduce the time for data analysis will be presented. A prototype system to analyze and align the sequence data with algorithms designed to be consistent with current base calling practices in the forensic community will be presented.

Peter R. De Forest Department of Sciences, John Jay College of Criminal Justice, CUNY, New York, NY

The meeting theme of "bridging" is an appropriate one. The meeting venue gives it extra symbolism. Two of the World's major bridges are here. They span two dimensions. One, a single span suspension bridge, once the longest in the world, extends north-south over the Golden Gate entrance to San Francisco Bay connecting San Francisco to Marin County and points north. The other, a combination double-span suspension bridge connecting to a cantilevered section east of Yerba Buena Island, extends generally east-west across the San Francisco Bay. Both bridges are well over sixty years old and are essential to the Bay Area economy. The north-south and east-west directions spanned by these bridges represent two dimensions. Figurative bridges spanning two dimensions are also needed in criminalistics. As is the case in the literal example, a conceptual bridge ties together and unifies separate or scattered entities. To fully exploit the physical evidence in any but the simplest cases there is certainly a need for a bridge spanning the wide range of disciplines within criminalistics. The need for spanning disciplines is really the need to span a wide range of physical evidence problems that may be encountered in a given case. Practitioners in these disciplines cannot operate in complete isolation. The other dimension to be spanned can be thought of as time. This time span is the life of the case. Alternatively this dimension can be thought of as spanning the stages in case development. It extends from "front end" assessment during the initial crime scene investigation to the "back end" interpretation designed to assist with the adjudication of the case. Between these is the continuum that includes evidence recognition, laboratory assessment, prioritization, analysis, interpretation, integration, reporting, explanation, and expert testimony. There is a need for at least one scientifically trained and experienced mind to have oversight over the entire physical evidence situation from beginning to end. This scientist must serve as a bridge for physical evidence and physical evidence derived data from the Crime Scene to the Courtroom. Bridging in the other dimension, across disciplines, is also essential simultaneously. It is necessary for at least one generalist-criminalist to view the case as a whole rather than having a narrow focus of specific evidence items or tests. It is also better if more than one scientist has this overview so that a sharing of ideas can take place as the approach to the case is being designed and executed. Case examples will be used. Most will describe situations where bridging was underutilized or absent initially, but where, fortuitously, some aspects of bridging could be taken advantage of later and the case solved. These cases are useful for illustrating the need for bridging. In effect there is a built-in negative control. In each the wrong conclusion or no conclusion was reached initially. It was only after the totality of the physical evidence picture was considered by a generalist-criminalist that the case was solved months or years later. Of course, in many cases where conceptual bridging is not taken advantage of, there is no "second bite at the apple" and the opportunity is lost. There is no substitute for doing it right the first time.

Peter J. Diaczuk., Zvi Herschman, Peter A. Pizzola and Peter R. De Forest Department of Sciences, John Jay College of Criminal Justice, CUNY, New York, NY

Casework experience and theoretical considerations have suggested that widely accepted beliefs about bloodspatter resulting from gunshot wounds are simplistic and often simply wrong. We have observed "expert" testimony founded on such false premises, which may have led to miscarriages of justice. These concerns prompted our research. In an earlier paper by some of us, it was our contention that bullets passing through portions of tissue supplied solely by capillaries would produce no bloodspatter. We asserted that special conditions needed to be satisfied in order for either forward spatter or back spatter to result from a bullet wound. Although we were able to cite casework examples and discuss theoretical reasons why no bloodspatter should result in many circumstances, we lacked a good experimental model to demonstrate our thesis. It was clear that models such as blood-soaked sponges, which have been used by others, are grossly inappropriate as a simulation of capillary supplied tissue. Saturated open-cell foam has a vastly higher blood loading on a volumetric basis than would capillary supplied tissue. Further, the blood would be much more finely divided in uninjured living tissue. In our earlier experiments we attempted to use models based on blood-perfused sheep brains and ballistic gelatin containing numerous small blood-filled channels. For reasons to be discussed, these were not satisfactory. Our new model is based on the use of arrays of blood-filled hollow fibers comprising dialysis cartridges. Work with these cartridges has supported our thesis. This thesis will be illustrated with casework examples and experimental results.

Nicole Inacio & Jennifer Matsumoto Department of Justice, Bureau of Forensic Services, DNA Laboratory

September 11th, 2001 left many of us fearful for our futures. In our despair at a moment of great vulnerability it was the desperation to help out in any way we could that brought us hope and strength. On January 5th 2002, a small group of volunteers from the Department of Justice DNA Laboratory in Berkeley set out to do just this. Without knowing how well we would be received, we braved the unmarked roads of New Jersey, and set forth to find Fresh Kills Landfill to aid with recovery efforts. Finding the landfill, merely by smell alone, we were given the opportunity to help out as much as we could. We spent 8 long hours a day at the largest crime scene in the history of the United States shoveling mud, sifting through debris, and walking through mounds of rebar and twisted metal in search of remains and personal effects. For such arduous labor, those 8 hours were one of the most emotionally profound and rewarding moments of our lives. We were there to aid in the recovery of remains and personal effects to give some closure, some finality to families, while helping the heroes of September 11th take a brief moment of rest and to hear their stories.

Robert A. Jarzen Sacramento County District Attorney's Laboratory of Forensic Services

The Sacramento County District Attorney's Laboratory of Forensic Services is pleased to announce that the development of the Trace Evidence Resource Center is currently underway. Funded through California's Office of Criminal Justice Planning - Local Forensic Laboratory Improvement Program, the Trace Evidence Resource Center will support and maintain sophisticated analytical instrumentation to resolve complex trace evidence cases that may otherwise go unsolved. Over the next 18 months, the Trace Evidence Resource Center will install advanced analytical instrumentation, such as: Inductively Coupled Plasma Mass Spectrometer (ICP/MS), Scanning Electron Microscope with Energy Dispersive Spectrometer (SEM/EDS), Liquid Chromatography Tandem Mass Spectrometer (LC/MS), FT-Raman Spectrometer, UV-Vis Microspectrophotometer. The Center will develop cooperative working agreements with California state and local crime laboratories to provide assistance in the analysis and interpretation of trace evidence casework. As part of this service, the Center will provide a centralized repository of trace evidence reference and exemplar materials. A visiting scientist program for research and procedure validation is also being considered.

Lara Walker and Meghan Kinney California Department of Justice, Bureau of Forensic Services-Freedom Lab

This case presentation is a combination of observations and documentation via photographs of a face impression that was left in the hood of a truck suspected in a felony hit and run. Although an impression in a hood is not necessarily remarkable, the detail observed and documented clearly demonstrates that the suspect did not hit a deer as he claimed. We will discuss the examination of the subject vehicle and the follow- up laboratory trace work and headlamp examination that was performed. The evidence presented in the criminal trial will be reviewed including an alternate theory proposed by the defense. We will show that seemingly powerful physical evidence does not always guarantee a conviction.

Mary Likins Northern California Innocence Project, Santa Clara University School of Law

Participant will understand the history of Innocence Projects in general and NCIP in particular. Participant will be able to relate the criteria for NCIP cases, the investigation process, and the types of physical evidence involved. Participant will also gain understanding of the current and proposed legislation in Sacramento related to post-conviction DNA testing, appointment of counsel, preservation of evidence and discovery. Participant will also hear anecdotal information about cases in which the evaluation of physical evidence has become an important aspect of the case, either to support or refute the inmate's claim of innocence. As Innocence Project cases progress, the need for experienced multi-discipline criminalists becomes ever more clear. The majority of cases handled by NCIP are not simply "up-or-down DNA" cases, but may require evaluation from crime scene investigators, coroners, trace evidence experts and others skilled in not simply performing tests but assessing the relevance of the material tested as well as the results.

Tom Malzbender, Hewlett-Packard Laboratories; Susan Morton San Francisco Police Crime Lab

Polynomial Texture Mapping, or PTM, was developed by Tom Malzbender and Dan Gelb of the Hewlett-Packard Laboratories as a way to create critically clear images of surfaces. Its initial purpose was to enhance computer graphics, but it was soon used by archeologists to render eroded clay cuneiform tablets legible. Its success in doing that has inspired inquiries from other fields such as dermatology and engineering. Our purpose in this presentation is to describe the method to forensic scientists in hopes that more applications may be discovered.

Charlene Marie California Department of Justice - Santa Barbara

The victim "repeatedly screamed for help while her husband allegedly beat her to death". She died of "blunt force trauma to her head and asphyxiation." Both the victim and her attacker were bleeding. This presentation is a reconstruction of the bloodstain evidence used to show the various locations of the attack; and, what could and what could not be said about the struggle. Her assailant was known; the pivotal issue was whether this was a case of 1st degree, 2nd degree or manslaughter.

Gerald R. McMenamin California State University, Fresno

To demonstrate that the measurement of variation in written language strengthens the descriptive analysis of style in cases of questioned authorship, and to respond to recent judicial specifications that evidence in forensic stylistics reflect appropriate quantification of language data in questioned authorship cases.

Susan Morton San Francisco Police Department

In the summer of 2001, Robert Nawi was tried and convicted of the first degree murder of Virginia Lowery in San Francisco. It was a case that would never have been solved much less brought to trial without three branches of forensic science. A fingerprint was lifted at the scene from a water heater near the body. At autopsy, the Medical Examiner noted and preserved foreign tissue under Mrs. Lowery's fingernails. The fingerprint was searched through AFIS databases, but no match was made. Ten years after the murder, a repeat AFIS search did result in a match to the fingerprint of one Robert Nawi. Mr. Nawi was an associate of William Lowery, but was not so lucky in his dealings with the law. He had pled guilty in federal court to drug dealing and had spent most of the intervening ten years in federal prison. Robert Nawi's DNA matched that under the victim's fingernails. Based on that evidence and the fingerprint identification, Nawi was brought to trial on murder charges. His defense was an alibi. He supplied affidavits from William Lowery, now living in Mexico and in poor health, and other then associates to the effect that he was in Mexico with them at the time of the murder and for some time before and after. The defense then attacked the fingerprint and DNA evidence. None of the forensic evidence was novel or cutting edge. Alone, any one would not have carried the day. But in combination, they made a very strong case. At least the jury thought so.

Norah Rudin Forensic DNA Consulting

Biological evidence, historically analyzed using conventional serology, now using DNA technology, has come to be treated as an entity separate from the rest of forensic science. It has been sequestered as a tool solely limited to answering the "who?' question that is often critical to case investigation. This limits our understanding of biological evidence and how it integrates into the case as a whole. Initially, biological evidence is transfer evidence, no different than any other trace evidence. From this perspective, the same questions can be asked of biological evidence as are typically asked of classical trace evidence: "what? where? how? when?" In fact, these questions have returned to the forefront with a vengeance as the "who" becomes almost indisputable in many cases. The Forensic Paradigm will be used as a framework to show how the interpretation of biological evidence can be reintegrated with other forensic disciplines. Practical applications will be illustrated using case examples. This type of unifying approach advances our understanding of criminalistics as coherent discipline with fundamental principles.

Sudhir K. Sinha*, Gina M. Pineda, Sharon Williams, Robin DeVille, and Alison Fleming ReliaGene Technologies, Inc.

Mitochondrial DNA analysis has been utilized in many forensic cases for the identification of victims whose bodies have been subjected to harsh and severe environments such as extremely high temperatures, humidity, and burials in shallow graves. This environment, which leads to the extreme degradation of the DNA from the bodies, has made mtDNA the method of choice for analysis of these cases. Close to 200 individuals in Panama have been victimized during the decades of military dictatorship that ended in 1989 with the ousting of Manuel Noriega. Today, Panama is a democracy, and the country has formed a "Comision de la Verdad" (a "Truth Commission") to locate and identify the missing bodies. Extensive efforts in Panama involving canine and anthropology disciplines led to the excavation of numerous remains. Sample processing techniques used to overcome PCR inhibitors will also be discussed. These include the use of an ultrasonic cleaner to remove any contaminating surface debris, the determination of DNA quantities and any protein contaminants or inhibitors in the sample, and the subsequent Qiagen column cleaning procedure used to rid the samples of inhibitors. Alternate PCR approaches, such as the use of mini primer sets, will be presented. Success rates of the remains will be discussed as well as rare mtDNA sequences that had never been observed in the available database beforehand. Identical sequences between remains obtained from different physical locations, and the implications of this finding, will be analyzed. Positive identifications made to date will also be discussed.

Theresa Spear* and Neda Khoshkebari California Criminalistics Institute

In an effort to ascertain the feasibility of successfully analyzing old biological samples from cases that could be more than 10 years old in the "Cold Hit" program (a program funded to find and type biological evidence from suspectless cases in California), we requested that crime laboratories in California send us old biological samples they could spare and provide a description of each sample and how it had been stored. The samples that were analyzed in this study included blood, semen, and saliva samples and included a sample that was 49 years old. The samples were described as being held in a variety of conditions: frozen, refrigerated, room temperature and outdoors. Samples obtained from crime labs in California were extracted, quantified and amplified for Profiler Plus® loci. On most samples, it was still possible to obtain positive body fluid identification test results using various body fluid identification tests (e.g. presumptive tests for blood and semen, immunological tests and amylase diffusion). Two semen stains (made in 1989 and 1975) did not give a positive AP or P30 test. A few of the very old bloodstains (25 years or older) gave weak or negative Hemastix test results. The most problematic test (i.e. the test producing the greatest number of false negative test results) in this study was the species test. The failure to obtain positive species results on some of the bloodstains tested was likely a result of the fact that these samples were very old and thus very insoluble. It is possible that had the stain extraction time been extended, more positive results would have been obtained.

Kenton S. Wong San Mateo County Sheriff's Forensic Laboratory

Two over-the-counter PASD were examined for their ability to accurately determine an individual's blood alcohol concentration (BAC). The crime of driving under the influence (DUI) of alcohol and its effects on society have received much negative notoriety in recent years. Without a doubt, the influence of groups such as Mothers Against Drunk Driving (MADD) have had a profound effect on placing the spotlight on the far-reaching ill effects of DUI in our local communities, as well as on a national level. As a partial result of this, legal limits of BAC have decreased considerably over the years. In the past, the only information provided and available for individuals to estimate their BAC was by using charts supplied by the Department of Motor Vehicles; however, this information was very difficult for the average individual to utilize. In response, several over-the-counter PASD have recently entered the marketplace purportedly allowing an individual to accurately monitor and determine their BAC by utilizing such devices. A number of these devices can readily be obtained from local drug stores at a meager price of only $0.99 to catalog mail-order suppliers or mall chain stores such as The Sharper Image for less than $100.00. Two of these devices were evaluated for their ability to accurately determine an individual's BAC.

Francis Woo San Francisco Police Department

GC/MS is widely used by most modern forensic laboratories for drug analyses. Since the decomposition pathways of analytes are thermodynamically controlled, certain fragment ions (m/e) are predictable. For molecules with similar structural isomers, it is important for the chemist to be able to explain and identify the variations in fragments. Both 3,4-MDMA and 2,3-MDMA produce similar mass spectra. A mechanistic decomposition approach is employed to explain the subtle differences to definitively characterize and differentiate each compound. Furthermore, a microcrystalloscopic method for differentiating 2,3 and 3,4 analogs of MDA and MDMA is also presented.

Kim Bogard*, Meri Bozzini, Danielle Blecha, Yasser Daoudi, Craig Leibelt, Rhonda Roby, Ariana Wheaton, and Farideh Shadravan

Forensic DNA laboratories using the Applied Biosystems' ABI PRISM® 3100 Genetic Analyzer coupled with four-dye and/or five-dye AmpFlSTR® products can increase their throughput capabilities for short tandem repeat analyses. The four-dye technology, using three (3) dyes to label DNA fragments and a fourth dye for the internal size standard, and the five-dye technology, using four (4) dyes to label DNA fragments and a fifth dye for the internal size standard, are both wellestablished, reliable techniques for forensic DNA fragment analysis. PCR products from four-dye (e.g., AmpFlSTR® Profiler Plus™ PCR Amplification Kit) and five-dye (e.g., AmpFlSTR® Identifiler™ PCR Amplification Kit) systems have been evaluated on three (3) 3100 Genetic Analyzers. The experiments performed include an evaluation of accuracy, precision, reproducibility, sensitivity, and mixture studies. Additionally, a new data collection software package was evaluated. A summary of these experiments will be presented as well as the running parameters.

Brian Burritt, San Diego Police Department

An excel spreadsheet that will calculate STR profile frequencies from any number of population studies has been developed. Currently, the program contains data from more than 130 population studies that contain, at a minimum, the nine Profiler Plus STR loci. The program was developed to aid the criminalist in quickly finding available population studies and performing DNA profile frequencies calculations with these studies. The compilation may be used to help demonstrate the worldwide use of STR testing. Additionally, the program can be useful as a research tool. Studies were carried out that addressed several assumptions that may be made regarding commonly used population studies. One assumption investigated, which is alluded to in NRC II, is that a DNA profile frequency estimate is likely within one order of magnitude from the "true" value. Another assumption investigated is that the presentation of Caucasian, African American, and Hispanic population frequencies gives a wide range of values that encompass the population frequencies of many of the world's populations. Population data generated with this program supports both of these assumptions. Additional population studies will be added as they become available. Copies of the program may be obtained from the presenter by email (k9b@pd.sannet.gov).

Mindy K. Crow

Bone samples have traditionally been problematic in terms of extraction of amplifiable DNA. The presence of PCR inhibitors, as well as significantly degraded DNA, are common problems observed with forensic bone samples. Three methods including the Dynal Dynabeads DNA DIRECT, sodium acetate/isopropanol, and QIAGEN QIAamp were compared to the commonly used phenol/chloroform extraction method in terms of their ability to extract a sufficient yield of amplifiable DNA. After DNA extraction and quantitation, the samples were carried through the protocols for the Amplitype PM + DQA1 PCR Amplification and Typing Kit. The sodium acetate/isopropanol extraction method was consistently successful at extracting DNA that amplified at all six genetic loci. The QIAGEN QIAamp method was also more effective than the phenol/chloroform method but provided slightly less consistent results than the sodium acetate/isopropanol method.

Lisa Lane, Promega Corporation

The DNA IQ™ System is a DNA isolation and quantitation system designed specifically for the forensic and paternity community. This system employs a novel technology with magnetic particles to prepare clean samples for short tandem repeat (STR) analysis easily and efficiently. The DNA IQ™ System can be used to extract DNA from stains, differential extracted sperm and epithelial fractions or liquid samples such as blood or resulting DNA solutions. The system is designed for casework database and paternity applications. For database samples the eluted DNA is delivered at about 1ng/μl eliminating the need to quantitate. The DNA IQ™ System for small casework samples includes two steps. For biological material on solid supports, the first step provides an easy, rapid, efficient and almost universal stain extraction method. This step is unnecessary for liquid samples. The second step uses a specific magnetic resin that purifies the DNA without requiring extensive washing to remove the lysis reagent. DNA is purified using a proprietary magnetic particle that removes virtually all PCR amplification inhibitors including dyes found in blue or black denim, soil and black leather. In addition, both the sperm and epithelial fractions generated from rape kits can be input directly into the purification process. To further simplify this process, the DNA IQ™ System has been tested with all Promega products and has been adapted to work on automated platforms. For paternity and database samples, the magnetic Resin captures a consistent amount of DNA. The Resin has a defined DNA capacity in the presence of excess DNA and will only bind a specific amount of DNA. This property is used to isolate approximately 100ng of DNA from a range of liquid blood, stains or swabs. The DNA is eluted into 100μl of Elution Buffer to give a DNA concentration of approximately 1ng/μl. As a result, the researcher can bypass the quantitation step typically necessary in other purification procedures.

Theresa Spear*, Sharyl Barney, Ashlie Silva and Neda Khoshkebari California Criminalistics Institute

This two-part study was designed to look at the impact of reagents used to characterize body fluid stains and reagents used to develop fingerprints on the ability to obtain PCR-based DNA typing results. Part I - Impact of Body Fluid Identification Reagents, Part II - Impact of Fingerprint Reagents In summary, clear-cut typing results were obtained for all 14 PCR-based markers from the DNA extracted from the bloodstains treated with each of these 6 reagents. Since the bloodstains in this study were made on absorbent, cotton swabs, this study did not examine the effect of the mechanical manipulations on the possible loss of sample. This would be an important factor consider when dealing with a limited amount of blood on a non-absorbent surface (e.g. plastic/glass). Any significant loss of sample amount can negatively impact typing results.

Mark D. Timken California DOJ Berkeley DNA Lab

In the California DOJ Offender Databank program, reference samples are currently obtained by collecting whole blood from qualifying offenders. Although blood is an excellent source of reference DNA, its collection is invasive and expensive, and its handling exposes collectors and analysts to potential bloodborne pathogens. As an alternative that minimizes these problems, we are developing protocols that use buccal swabs as the source of reference DNA. Automated protocols have been developed using a Tecan Genesis 150 Robotic Workstation for magnetic-bead-based and FTA®-paper extraction methods. Results from these protocols will be compared, mainly with respect to their first-pass genotyping success rates.