[Final Version]

DNA Forensic Evidence

By Ryan Turner Rachel Kosa Bruce Kubu

May 2002

POLICE EXECUTIVE RESEARCH FORUM 1120 Connecticut Avenue, N.W., Suite 930, Washington, DC 20036

Contents
Introduction..................................................................................... 1 Background..................................................................................... 2
DNA Description............................................................................................ 2 DNA Analysis................................................................................................. 3

Collecting Evidence ....................................................................... 3

Basic Considerations.................................................................................... 5

Appendix ....................................................................................... 14
Restriction Fragment Length Polymorphism (RFLP) ............................... 14 Polymerase Chain Reaction (PCR) ............................................................ 16 Mitochondrial DNA ...................................................................................... 17

Resources ..................................................................................... 18
National Organizations and Training Resources (fee and non-fee)........ 18 Model State Crime Lab Programs .............................................................. 18 Contacts ....................................................................................................... 18 Reviewer ...................................................................................................... 19 Selected References ................................................................................... 19

DNA FORENSIC EVIDENCE

Introduction
In recent years, several high-profile homicide cases have used DNA analysis as evidence. Famous examples include the 1995 O. J. Simpson trial and the investigation into the 1993 death of Deputy White House Counsel Vince Foster. But DNA evidence is no longer used solely in high-profile or especially difficult homicides. In 1996, it was used in more than 17,000 court cases. The FBI has developed the Combined DNA Index System (CODIS), a software system used on the local, state, and national level, to link DNA testing results from unsolved crimes. The CODIS system is very similar to the Automated Fingerprint Identification System (AFIS) database. In the same manner that fingerprints can be run through AFIS to identify an offender, DNA profiles from a crime scene may also be entered into CODIS to search for a suspect or to link the offender to another crime scene (What Every Law Enforcement Officer Should Know About, National Institute of Justice). As of summer 1998, CODIS had been used to aid 583 investigations, had yielded 233 case-tocase hits, linking together multiple crimes, and had 182 case-to-offender hits. In October 1998, the FBI introduced the National DNA Index System (NDIS), which will allow laboratories around the United States to exchange and compare DNA profiles electronically. As of October 1998, CODIS was being used by 94 laboratories in 41 states and the District of Columbia. NDIS will allow labs to link DNA evidence from unsolved crimes. While DNA evidence may be the crucial piece of evidence that results in a conviction, it may also exonerate innocent suspects. This ability helps police satisfy their duty to present exculpatory evidence. It is estimated that DNA testing has excluded the named suspect in approximately one-third of cases where DNA analysis was performed. DNA testing has also been used to exonerate people wrongly convicted of a crime. As of 1996, at least 28 individuals convicted of crimes, and serving time in prison, were released due to DNA analysis of evidence after the trial.

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

Background
The goal of DNA analysisor genetic typingin forensic investigation is to identify potential donors of biological stains (blood and other body fluids) found at the crime scene or elsewhere. Even though it is a more precise analytical tool than other types of forensic evidence, DNA evidence is still prone to technical uncertainties and skepticism. Once its potential to aid homicide investigations is fully understood, crime scene personnel and investigators can be more effective in collecting, handling, and preserving DNA evidence.

DNA Description
Deoxyribonucleic acid (DNA) is the molecular basis of inheritance, acting as a blueprint for each individual. DNA is inherited from both parents, passing genetic traits from one generation to the next. It is found in every cell of the body that has a nucleus, including white blood cells, sperm, and exterior skin cells. Although hair shafts do not contain DNA, hair roots may be used for analysis since they contain cells with nuclei. Each cell carries exactly the same genetic information. DNA exists in the form of microscopic strands called chromosomes, which resemble very long, twisted ladders. Each strand is made up of pairs of units called bases. A gene is a section of DNA containing a specifically ordered set of thousands of base pairs. Genes represent unique molecular codes for protein building blocks, which determine traits such as eye color and blood type. The position a gene occupies along the DNA thread is its locus; every individual has one gene from each parent at a given locus. Each locus can contain as many as 100 different gene forms. These alternate forms are called alleles. Different allele combinations result in variations of a trait, such as eye color. Forensic analysts are interested in both the potential number of allele combinations and the relative frequency with which certain combinations appear within a population. If a geneticist or forensic scientist knows the frequency of a certain allele combination, then he or she can calculate the probability two people in that population would have the same allele combination.
POLICE EXECUTIVE RESEARCH FORUM 2

DNA FORENSIC EVIDENCE

DNA Analysis
The entire complement of a persons DNA (called the genome) consists of approximately 3 billion base pairs, of which approximately 3 million pairs (0.1 percent) differ from person to person. DNA typing seeks to identify and isolate discrete portions of these variable DNA strands to compare one sample with another for the presence of similar fragments. Because those fragments are so specific to individuals, DNA evidence found at a crime scene can provide a probability whether a specific person deposited their genetic material at that location. It is especially useful in cases where there is contact or an exchange of bodily fluids between the victim and the assailant, and particularly when sexual assault, rape, and sodomy are involved with another crime, like murder. DNA evidence can only corroborate or disprove an investigators other evidence. A lab can determine the genetic profile of a biological stain only when appropriate samples from the victim and suspect are submitted for comparison and the stain is large enough to extract DNA for analysis. The two genetic profiling techniques employed are restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR) (for a description of each of these techniques, refer to Appendix A).

Collecting Evidence
The most important aspect of DNA evidence collection is crime scene preservation and maintenance because the integrity of all potential evidence is at stake. Making an effort to collect the best possible evidence at the scene of a crime can aid in case closure and convictions at trial. Because DNA testing can also yield exculpatory evidence, however, old cases might well be reopened. If evidence is documented, collected, and stored properly, it can be presented to a judge or jury several years from the time of the criminal act. Police are the first responders to most crime scenes and are responsible for handling evidence before it reaches the crime lab. The repercussions of mishandled DNA evidence are significant:

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

Poor specimen quality can result in degraded DNA or relatively short DNA fragments, which are potentially difficult to analyze. Poor collection of trace evidence can cause unreliable forensic lab work or inconclusive results, which in turn necessitates the use of less reliable evidence (circumstantial evidence or eyewitness testimony) or less tangible evidence (lab reports, case notes, and photographs) during an investigation or trial.

The impact on a criminal trial can be reasonable doubt in the mind of the judge or juror or wrongful convictions in cases using DNA evidence.

Law enforcement personnel cannot take physical evidence at a crime scene for granted. Unless it is properly collected and preserved, and the chain of custody is maintained, even what is seemingly the most damaging evidence may be doubted by the judge or juror. The most important aspect of evidence collection and preservation is protecting the crime scene from the time the first officer or responder arrives until the last piece of evidence has been noted and collected without being contaminated. Only people responsible for the crime investigation should be present. Any other individuals or unescorted personnel can introduce contamination or disrupt the crime scene. The area can be secured using ropes, tape, cones, barricades, or even additional officers. A crime scene log or sign-in sheet should also be used to record who enters the area and when. In addition, all crime scene personnel must wear protective clothing, hairnets, gloves, paper boots, masks, and eye protection. Although some localities house criminalists in police departments, others may only have one or a few officers trained to handle evidence properly during an investigation and during and after a trial. No agency has every type of specialistserologist, criminalist, blood pattern analystavailable from among its ranks. Agencies should develop a working relationship with specialists before they are needed in an actual case, and an up-todate directory of specialists should be maintained for use by investigators. Specialists should be consulted regularly to determine the best way to conduct planning, operations, and follow-up activity jointly.
POLICE EXECUTIVE RESEARCH FORUM 4

DNA FORENSIC EVIDENCE

There can never be too much documentation of physical evidence. Investigators and crime scene personnel should thoroughly document every aspect of the crime scene investigation, from the initial walk-through to the securing of the collected evidence, by using notes, videotape, photographs, sketches, and chain-of-custody forms. Thorough documentation can assist investigators who need to revisit the case in the future. Only after all evidence is properly documented should any evidence retrieval begin. Many factors contribute to inadequate collection of potential DNA evidence, later compromising its use: acts by the offender that render unlikely the deposit or recovery of trace evidence contaminants such as bacteria, victim and suspect specimen mixing, and nonhuman DNA sources lack of knowledge about which stains yield the most useful information, and poor reference samples of those stains lack of communication between law enforcement personnel and forensic specialists, lab and evidence technicians, and medical examiners misconduct on the part of law enforcement and scientific experts (such as misplacing or prematurely destroying evidence, or lying) There is only one chance to perform the collection of DNA evidence properly. Crime scene personnel should therefore take a slow and methodical approach to collecting and preserving it. Unless the evidence is in danger of being destroyed or compromised, the best searches are typically cautious, difficult, and time-consuming.

Basic Considerations
All evidence must be delivered to the examining lab as soon as possible. Most items of evidence should be collected in clean, unused paper containers (packets, envelopes, and bags) to prevent contamination by other evidence. Plastic containers should never be used. Each container should have the collecting persons initials; the date and time the evidence was collected; a complete de-

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

scription of the evidence and where it was found; the investigating agencys name; and the file number. Control samples (those not taken from evidence) should be air-dried, packaged in paper, and then frozen (not refrigerated) until used by the lab. Items at the crime scene should be examined to determine if there is any loose trace evidence (hairs, fibers, paint chips, etc.) that may be lost in transportation. This loose evidence should be collected in a paper packet and placed in an envelope, with a description of the trace evidence and its source. Under no circumstances should evidence containing moisture be sealed in plastic or paper containers for more than two hours. Moisture allows for the growth of microorganisms that can destroy or alter evidence. The chart on the following page details some common procedures and considerations to remember when collecting and storing DNA evidence for processing by the analyzing laboratory.

Evidence Type Blood (see note at end of table) Form Dried stains (small items)

Collection Method If possible, wrap the item in clean paper, place the article in a brown paper bag or box, and seal and label container. Send the whole stained object to the laboratory after labeling and packaging. Do not mix dried stains; place each in a separate envelope. Never attempt to wipe dried stains from an object using a moistened cloth or paper. Do not attempt to remove dried blood stains from clothing.

Risks

Special Considerations

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

Evidence Type Blood Form Dried stains (large items)

Collection Method Preferred Method: Cover the stained area with clean paper and seal the edges down with tape to prevent loss or contamination.

Risks More work for the serologist: bulky items require more storage space.

Special Considerations Requires a minimal amount of interaction with the bloodstains by the investigator and allows the serologist to make the decisions involved in collecting the samples. Dilution and contamination potential minimized by eliminating the use of water as the collection medium. Dilution and contamination potential eliminated by not using water as the collection medium. Investigator has minimal interaction with the bloodstain, and evidence does not take up much storage space. A fairly easy technique in which the control sample is readily collected. Dilution and contamination potential minimized by eliminating the use of water as the collection medium. Requires little storage space.

Alternate Method #1: Cut out the part of the item with the bloodstain(s). A control sample should also be cut out if available. Both cuttings should go into separate paper envelopes.

Investigator must use discretion to determine which stains and controls to collect. Some materials are difficult to cut through.

Alternate Method #2: Use fingerprint tape to lift bloodstain. Place tape (do not touch sticky surface with bare hands) over bloodstain and surrounding negative control area. Rub non-sticky side of tape with a pencil eraser or blunt object to insure good contact between stain and tape. Lift the bloodstain like a fingerprint and place the tape on a vinyl acetate backing (paper backing makes the stain difficult to handle during analysis). The process can be repeated several times on the same stain if necessary. Label the stain and package in a paper envelope. Blood (continued) Dried stains (large items) Alternate Method #3: Scrape bloodstains into a paper packet with a clean, sharp instrument. Label packet and place in a paper envelope. Never use plastic containers, which create static charge that makes blood flakes scatter and stick to the container interior.

Investigator must decide which stains and controls to collect. Bloodstains do not lift well off certain surfaces.

Investigator must decide which stains to collect; when scraped, bloodstains break into small, difficultto-handle flakes; flakes are easily lost during the scraping process; some surfaces are not easy to scrape.

Dilution and contamination potential minimized by eliminating the use of water as the collection medium. Requires little storage space. [NOTE: Blood flakes can be scraped near the sticky side of some fingerprint tape. The static charge will cause the flakes to stick to the tape, and the tape can be placed on vinyl acetate.]

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

Evidence Type Form

Collection Method Alternate Method #4: Absorb stains onto long, number 8 white cotton threads moistened with distilled or deionized water. Threads should not be touched with bare hands. Threads are placed on bloodstain with a pair of clean forceps or a clean cotton swab and rolled on the stain until at least four threads are collected. Threads and swabs are air-dried and put into a paper packet and envelope. Threads may be placed in a plastic container for no more than two hours. Once in a secure location, the threads must be removed from the plastic and allowed to air dry. They may then be repackaged into a paper packet and placed in an envelope. Threads must also be taken from a negative control area, if available. Alternate Method #5: Absorb stains onto moistened x cotton squares, following the same procedure as with threads. The muslin must be boiled in distilled or deionized water and air-dried before use to remove interfering factors. Bare hands should not touch the muslin. Place small stained items in paper bag (or plastic bag to prevent contamination of other objects). In a secure spot, take item out of bag, and allow the evidence and bag to thoroughly air dry. Do not dry stained material by using heat or placing it in bright sunlight. Repackage in the original paper bag or, if necessary, a new paper bag. If a new paper bag is used, then the air-dried original container should be packaged with the item of evidence. Absorb the stain onto a 1 x 1 square of cotton muslin. Package it in paper (or plastic to prevent contamination of other objects). Bring it to a secured location, take it out of the container, and allow the cotton square and the container to thoroughly air dry. Do not dry stained material by using heat or placing it in

Risks Dilution and contamination potential is increased due to using water; investigator must use discretion as to which stains and controls to collect.

Special Considerations Stain is concentrated onto a relatively small surface area, requiring little storage space.

Dilution and contamination potential is increased due to using more water.

Stain is concentrated onto a relatively small surface area; easier to handle than threads; requires little storage space.

Blood (continued)

Wet stains (small items)

Evidence should be refrigerated or frozen immediately, then delivered to the laboratory as quickly as possible. Delays beyond 48 hours may increase the chances of decomposition. More work for the serologist; bulky items use more storage space.

Requires a minimal amount of interaction with the bloodstains by the investigator; allows the serologist to make the decisions involved in collecting the samples.

Blood

Wet stains (large items)

Evidence should be refrigerated or frozen immediately, then delivered to the laboratory as quickly as possible. Delays beyond 48 hours may increase the rate of decomposition and decrease the amount of recoverable DNA. Investigator

Requires little storage space; fairly easy technique to perform; stain is concentrated onto a relatively small surface area.

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

Evidence Type Form

Collection Method bright sunlight. Repackage in the original paper container or, if necessary, a new paper container. If a new paper container is used, then the airdried original container should be packaged with the cotton square. If possible, the investigator should also collect samples from unstained areas of the item for negative controls. Allow any stains to air dry. If damp, allow fabric to dry completely before packaging. Wrap in paper and place in paper, not plastic, bag. Label and package each stained item separately If victim shows evidence of sexual intercourse, use PERK. If necessary, oral, vaginal, or anal swabs should be taken from the victim. Swabs should be air dried under a fan or moving air source for at least one hour. Samples should be stored refrigerated. Do not freeze samples as freezing and thawing can rupture sperm cells.

Risks must decide which stains and controls to collect; investigator must have direct interaction with bloodstain.

Special Considerations

Semen and seminal stains

On fabric

Often found on clothing, blankets, and sheets.

On victim

The body begins breaking down the various components in seminal fluid through drainage, enzyme activity, pH, etc. Moisture in the swabs allows microorganisms to grow, which can destroy the evidentiary value of the swabs.

Take swabs as soon as possible. Reasonable attempts should be made to take evidence from all viable suspects (including family, friends, etc.). Evidence collected and subjected to testing may reveal results from biological material left by other consensual sexual partners unrelated to the offense investigated or other contact with victim by other individuals.

Saliva

Clothing

Hair

Wet or not completely dry With root sheath

Use sterile gauze pad or swabs; allow to air dry. Place in paper, not plastic, containers. Sources of saliva can include envelopes, bottles, cans, gum, food, etc. Hang articles in a room with adequate ventilation and allow to air dry. Label, roll in paper, then store in brown paper bag or box; seal and label container. Collect 15-20 representative hairs from the suspect. Place in paper packet and then in an envelope.

Handle fabrics as little as possible.

Hair

Withoutroot sheath

Collect 15-20 representative hairs from the suspect. Place in paper packet and then in an envelope.

If a root sheath is attached, DNA analysis using PCR technology can provide information on the likelihood that this hair came from a certain percentage of the population to which the suspect belongs. If there is no root sheath, microscopic analysis can reveal whether the hair has the same characteristics as the suspects hair. Addi-

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

Evidence Type Form

Collection Method

Risks

Special Considerations tionally, mtDNA testing may also be used.

Stain evidence on nonabsorbent materials

On materials such as plastic and metal, shifting the material from a cold to a warm environment may create condensation, destroying the forensic value of the sample. Samples must be packaged so the stain portion is protected. Keep evidence at room temperature and deliver to lab as quickly as possible.

Note: Blood evidence is generally more informative in cases where a suspect and victim were in contact or close proximity. Crime scene personnel should concentrate on collecting representative samples of the peripheral bloodstains, such as those that are away from the body and the main area of action, or blood spatter patterns that differ from the majority of the blood spatter patterns, all of which may provide useful investigative information. Luminol, when applied to even very dilute bloodstains, will cause them to glow in the dark. However, it has serious drawbacks as a presumptive test for the presence of blood. Luminol also gives false readings when it reacts with stains containing copper ions and compounds, iron compounds, cobalt ions, potassium permanganate (found in some dyes), hydrated sodium hypochlorite (bleach), ferricyanide, and plant peroxidases. Because luminol is water-based, it can cause latent impressions to smear. The investigator should first use an alternate light source, such as ultra violet (UV) light, to search for any traces of blood, semen, or saliva. Diluted blood often leaves a brownish stain where a person has tried to clean it. Blood also has a tendency to flow into floorboard cracks, into carpet padding, behind baseboards, etc. These items can then be collected and submitted to the crime lab for confirmatory testing. Comparison testing must be conducted between the genetic markers in the victims blood, the suspects blood, and reference samples found at the crime scene. All blood samples should be collected in a purpletop Vacutainer tube, stored in a refrigerator (not a freezer) at four degrees centigrade until delivered to the crime lab. Investigators should consult in advance with the examining lab to determine the preferred container for reference samples. Bloody clothing and other stained items are poor reference samples because their sources may be questionable, and various other factors may give misleading results.

POLICE EXECUTIVE RESEARCH FORUM

10

DNA FORENSIC EVIDENCE

The more hands that are involved with the evidence once it is out of police custody or proper channels, the less chance there is to recover the evidence intact if it is lost. After evidence has been presented at trial, it may be retained by the court or remitted to the investigating agency, state police, or medical examiners office. Courts and medical examiners or labs have limited space and therefore are sometimes unwilling to hold onto the evidence. If a case is reopened, it is better to have quick access to the evidence and information. Keeping records and documentation is a natural task for the office that gathered the evidence, so this is where evidence should ultimately remain, namely in police custody. It is therefore important that a chain of custody not only be developed but well documented. A chain of custody form should record all transfers of a DNA evidence sample, from collection at the crime scene to possession by the court. This documentation is kept to ensure the integrity of the chain of custody with respect to the evidence, from the moment of collection through presentation in court. The form should include the following information: case number description of the evidence and its container specific recovery location time and date of the evidence names of persons who collected it notation of whether container was sealed upon transfer to another individual names of persons who have received evidence and the time and date of those transfers final state of the evidence

The form should accompany the evidence all the way to its final destination, and a copy should be kept in the case folder.

POLICE EXECUTIVE RESEARCH FORUM

11

DNA FORENSIC EVIDENCE

Conclusion
DNA analysis gives police investigators a very powerful tool in the fight against crime. Forensic use of DNA is growing, and findings are both convicting the guilty and exonerating the innocent. To make good use of this capability, however, police officers and investigators need to become skilled in identifying, collecting, transporting, storing, and tracking biological specimens. Both scientific and legal strictures require that such evidence be handled carefully and correctly.

POLICE EXECUTIVE RESEARCH FORUM

12

DNA FORENSIC EVIDENCE

Appendix
RFLP The lengths of various DNA fragments along chromosomes differ among individuals. This phenomenon is known as polymorphism. Restriction fragment length polymorphism (RFLP) analysis permits police to identify or eliminate an individual as the contributor of a particular stain containing blood, seminal fluid (with sperm), vaginal fluid, skin tissue, or hair samples that include the root. RFLP analysis consists of seven basic steps performed in a lab: Extraction. DNA is extracted from the sample. Fragmentation. The resulting DNA molecules are subjected to enzymes that cut the DNA strands into smaller fragments at very specific base pair sequences. Electrophoresis. DNA fragments are placed in a semi-solid gel to which an electric current is applied. The DNA molecules, which carry a natural negative electronic charge, migrate across the gel towards a positively charged electrode, and the smaller fragments move faster than the larger ones. When the electrophoresis is completed, the fragments are arranged on the gel according to their lengths. Southern blot transfer. Fragments are lifted away from the gel and onto a nylon membrane via capillary action between a buffer solution and a layer of filter paper. The fragmented strands are then pulled apart at the base pairs. Hybridization. The resulting fragments on the nylon membrane represent an individuals entire DNA complement or blueprint. To identify the alleles necessary for comparison, lab technicians use either radioactive or chemical luminescent techniques. These involve adding radioactively labeled isotopes (P32) or enzyme labeled probes (short segments of DNA with base pair sequences complementary to the sample DNA) which seek out and attach themselves to the complementary sections of the sample DNA. Autoradiography. The treated membrane is then placed on a sheet of x-ray film. The radioactive or chemically treated probes emit either radioactivity or light, respectively, which in turn develops the film and reveals the location of the alleles

POLICE EXECUTIVE RESEARCH FORUM

13

DNA FORENSIC EVIDENCE

in the form of bands at different positions along the film. This print is known as an autoradiograph (or autorad) and provides a stable and permanent record of the DNA profile. Interpretation. Interpretation of the suspects and victims DNA prints involves comparing a sample of known origin with forensic specimens collected at the crime scene. A match is declared when all bands appearing in one test sample appear in the other. If the DNA profile from the evidence sample and that of the suspect match, they may have come from the same person, or they might represent a coincidental match between two persons who happen to share the profile. A match is more difficult to declare than an exclusion (non-match) because potentially corresponding bands in the two prints may not correspond perfectly, due to a number of conditions. The initial comparison is done with the naked eye. Computerized scanning and comparison of prints is used when an eye assessment is too close to call. Because the entire population cannot be tested to assess frequency, estimates derived from population genetics are used to determine the significance of the match, namely the likelihood that two, non-related persons coincidentally possess this identical combination of bands. After analysis, the probe is washed off the membrane, and the entire process is repeated for four or more loci. The number of tests that can be performed on a sample is limited by the risk of removing some of the DNA fragments, which could render the bands on the autorad weak or invisible. Originally, RFLP DNA analysis was given the name DNA fingerprinting, implying that each person has a unique DNA fingerprint. An expert may testify that a particular fingerprint belonged to a certain individual. However, up until 1997, the courts ruled that DNA results could only be given in statistical terms and that an expert could not testify that a DNA sample came from a specific individual. He or she could only testify as to the DNA profile of the individual and to the statistically estimated expected frequency of that profile in the given population (Caucasian, African American, Latino, Indian, etc.). In 1997, however, the FBI established a policy to guide testimony that may be given re-

POLICE EXECUTIVE RESEARCH FORUM

14

DNA FORENSIC EVIDENCE

garding a genetic sample. The probability that a DNA sample is that of the suspect is often described in terms of a random match, or the likelihood that a random person in the population would have the same DNA profile as the suspect. The FBI now considers a definitive identification to be declared within a reasonable amount of scientific certainty when the probability exceeds one in 260 billion. PCR Polymerase chain reaction (PCR), unlike RFLP, permits the analysis of extremely small amounts of DNA, thus providing a typing technique for samples too small to be used with other approaches. Through a repeated process of heating, cooling, and replicating enzyme exposure, the quantity of DNA can quickly increase by several million times. The PCR test uses an allele-specific probe to seek out specific polymorphic segments (alleles) and exponentially synthesizes exact copies of those sequences. Although RFLP analysis can more uniquely discriminate between individuals with the same number of probes or loci, it requires greater sample amounts than might be available. Blood evidence must be at least the size of a quarter, semen samples at least the size of a dime. Samples must also be well preserved, concentrated in one spot (not spread out), and not degraded. Radiographic RFLP analysis can take approximately 10 to 12 weeks while, with chemi-luminescent techniques, a lab technician can do four to five probes in two weeks. PCR testing, by contrast, takes approximately one week. Only a small amount of evidence is needed for testing, so portions of the evidence can be saved for future testing. In addition, old, degraded, or diluted evidence can be tested. PCR testing involves three steps. First, the DNA is extracted from the item. Then, the DNA is replicated and the replicated or amplified DNA product is detected with a variety of methods. However, PCR testing can be susceptible to contamination, as the replication process can confuse the interpretation of typing results if contaminant DNA is replicated along with the evidence DNA. Since the officer collecting evidence does not usually know which test (PCR or RFLP) will be employed, care and caution must be exercised in the collection and storage of evidence from which DNA samples might be drawn.
POLICE EXECUTIVE RESEARCH FORUM 15

DNA FORENSIC EVIDENCE

Like RFLP testing, PCR can be utilized with different types of genetic markers. Initially, PCR testing was used with a reverse dot-blot system to provide information on the DQ alpha gene, as well as five other markers collectively referred to as the polymarker test. These six genetic sites, which could all be tested for at once, were visualized by the presence of a blue colored dot along a nylon strip that had been treated with probes from the markers being tested. More recently, with the FBIs selection of a set of 13 short tandem repeat (STR) markers for CODIS, most forensic laboratories that conduct PCR testing are moving towards STRs so they can participate in the shared information in this database. STRs also amplify or copy extracted DNA, but then the DNA is run on an electrophoretic gel similar to RFLP analysis (though here the fragment sizes are much smaller). The gel is then either stained with a silver solution to visualize the resultant bands, or detection can be automated by using pre-packaged probes and software to produce an electropherogram (a graph with peaks that represent the alleles present in a specific sample). Though the FBI selected 13 STRs for CODIS, many additional STRs exist and can be utilized. The more markers used, the more uniquely discriminating the result. Mitochondrial DNA Mitochondria are organelles, or small, membrane-bound sacs found within a cell, but they exist in the cytoplasm, outside the nucleus of the cell where other DNA is found. Mitochondria contain DNA (mtDNA) that, like DNA found in the nucleus, is identical in all of a persons body tissues. However, mtDNA differs because it is present in very high numbers. Further, it is transmitted only through the mother, so all children from one woman have identical mtDNA. Maternal transmission proves a disadvantage for forensic use, because siblings or other persons sharing a maternal relative have indistinguishable mtDNA. Mitochondrial DNA also has less discriminatory power than nuclear DNA. Mitochondrial DNA can be of great use in cases where the sample is degraded due to age or environmental conditions, or where only a very small sample is available. For example, it is possible to extract and isolate mtDNA from a single strand of hair. Unlike conventional RFLP or PCR analysis of nuclear DNA, mtDNA is found in the actual shaft of the hair. A degraded or small sample may not have enough nuclear DNA to analyze but

POLICE EXECUTIVE RESEARCH FORUM

16

DNA FORENSIC EVIDENCE

may still contain enough mtDNA for a PCR analysis. The Armed Forces DNA Identification Lab uses mtDNA to identify the remains of soldiers. Although there is lower discriminating power with mtDNA and it is a relatively new technique, it does have some applications in the courtroom. However, mtDNA has the potential to become a more powerful tool in criminal cases in the future.

POLICE EXECUTIVE RESEARCH FORUM

17

DNA FORENSIC EVIDENCE

Resources
National Organizations and Training Resources
National Association of Crime Lab Directors FBI/Quantico Various state training and education resources Francis Glessner Lee/Harvard Associates Death Investigation School, sponsored through the Chief Medical Examiners office in Baltimore

Model State Crime Lab Programs

ConnecticutDr. Henry Lee VirginiaDr. Paul Ferrara Floridacollege and university programs and coordinated programs with the Highway Patrol (for vehicular homicide)

Contacts
Dr. Paul Ferrara Director Virginia Division of Forensic Sciences Virginia Crime Lab One N. 14th St. Richmond, VA 23219 (804) 786-2281 http://www.state.va.us/~dcjs/forensic Richard Payne and Sgt. Dave Sargeant Metropolitan (D.C.) Police Department Crime Lab S. Capitol St. Washington, DC (202) 727-6716 George Schiro
POLICE EXECUTIVE RESEARCH FORUM 18

DNA FORENSIC EVIDENCE

Louisiana State Police Crime Laboratory P.O. Box 66614 Baton Rouge, LA 70896 e-mail: 75054.1502@compuserve.com

Reviewer
Ms. Anjali Swienton National Institute of Justice Office of Science and Technology 810 Seventh Street, NW Washington, DC 20531 (202) 307-0645

Selected References
Bedau, Hugo, and Michael Radelet. In Spite of Innocence. Northeast University Press, 1992. Bureau of Justice Statistics. Forensic DNA Analysis: Issues, June 1991. Clarke, Alan. Procedural Labyrinths and the Injustice of Death: A Critique of Death Penalty Habeas Corpus, Part Two, University of Richmond Law Review, Vol. 30, p. 303 (March 1996). Confronting the New Challenges of Scientific Evidence, Harvard Law Review, Vol. 108, p. 1481 (May 1995). Convicted by Juries, Exonerated by Science: Case Studies in the Use of DNA Evidence to Establish Innocence After Trial. Washington: U.S. Department of Justice, 1996. DNA Technology in Forensic Science. National Academy Press, 1992. Evidence Handling Guide, Los Angeles Department of Public Safety and Corrections, Office of State Police, Crime Laboratory. FBI Forensic Serology Course Manual. Federal Judicial Center. Reference Manual on Scientific Evidence. 1994. Fisher, Barry A. J., Arne Svensson, and Otto Wendell. Techniques of Crime Scene Investigation. New York: Elsevier, 1981. Gianelli, Paul. Chain of Custody and the Handling of Real Evidence, American Civil Law Review, Vol. 20, No. 4, pp. 527-568 (Spring 1983).
19

POLICE EXECUTIVE RESEARCH FORUM

DNA FORENSIC EVIDENCE

Giannelli, Paul. Criminal Discovery, Scientific Evidence, and DNA, Vanderbilt Law Review, Vol. 44, p. 791 (May 1991). Haglund, W. D., and M. H. Sorg, eds. Forensic Taphonomy: The Postmortem Fate of Human Remains. CRC Press, 1997. Jonakait, Randolph N. Forensic Science: The Need for Regulation, Harvard Journal of Law and Technology, Vol. 4, p. 109 (Spring 1991). Journal of Criminal Law and Criminology, Vol. 84, No. 1, 1993. Krent, Harold J. The Puzzling Boundary Between Criminal and Civil Retroactive Lawmaking, Georgetown Law Journal, Vol. 84, p. 2143 (June 1996). National Research Council, The Evaluation of Forensic DNA Evidence. National Academy Press, 1996. Nature, Nos. 314, 316, and 318, 1985. Oops! We Forgot to Put the Evidence in the Refrigerator: DNA Identification and the States Duty to Preserve Evidence, The John Marshall Law Review, Vol. 25, pp. 809-836 (1992). Pearsall, Anthony. DNA Printing: The Unexamined Witness in Criminal Trials, California Law Review, Vol. 77, p. 665 (May 1989). Rayford-Williams, Claudia, and Andreas V. Smith. How Technology Has Affected the Legal System: Its All in the Genes: The Application of DNA Fingerprinting in the Courtroom, Howard Law Journal, Vol. 34, p. 139 (1991). Renskers, Sally E. Trial by Certainty: Implications of Genetic DNA Fingerprints, Emory Law Journal, Vol. 39, p. 309 (Winter 1990). Scheck, Barry C. DNA and Daubert, Cardozo Law Review, Vol. 15, p. 1959 (April 1994). Schiro, George. Collection and Preservation of Evidence, What We DoLaw Enforcement Series, compiled by Captain Merril L. Boling, Jefferson Parish (Louisiana) Sheriffs Office, 1995. Weed, V. W., and J. W. Hicks. The Unrealized Potential of DNA Testing, National Institute of Justice Journal, December 1997, pp. 16-23. What Every Law Enforcement Officer Should Know About, National Commission on the Future of DNA Evidence, National Institute of Justice, 1999.
POLICE EXECUTIVE RESEARCH FORUM 20

DNA FORENSIC EVIDENCE

Wood, Paul, Stephen Passen, and Don McKinney. 17th Annual Survey of Arkansas Law: Criminal Procedure, University of Arkansas at Little Rock Law Journal, Vol. 17, p. 501, (Spring 1995).

POLICE EXECUTIVE RESEARCH FORUM

21