Forensic science Forensic science is the application of science to criminal and civil laws, mainly—on the criminal side— during criminal investigation, as governed by the legal standards of admissible evidence and criminal procedure. Forensic scientists collect, preserve, and analyze scientific evidence during the course of an investigation. While some forensic scientists travel to the scene of the crime to collect the evidence themselves, others occupy a laboratory role, performing analysis on objects brought to them by other individuals.” In addition to their laboratory role, forensic scientists testify as expert witnesses in both criminal and civil cases and can work for either the prosecution or the defence. While any field could technically be forensic, certain sections have developed over time to encompass the majority of forensically related cases.Polymerase Chain Reaction (PCR) 4 If there is only a small amount of DNA available for DNA Fingerprinting the amount of DNA is increased by using a technique called PCR. PCR is a method of DNA replication in a test tube. * Like All DNA Polymerases Taq polymerase can only add to the 3’ end of an existing nucleotide +A DNA primer that is complementary to the template is used to supply that 3’ end ®o @ + + | GEEAET GAA e a Double stranded DNA Nucleotides Taq polymerase Heat to denature Taq polymerase adds complementary nucleotides to each single strand to make double stranded DNA. 1 round of : ' VNTRs replication (variable Cmmper=a CTT o number GTCATGAA ~— tandem ar. identical DNA ll are formed. <— repeats) « After we isolate = the DNA and Figure 7-7a Biology: Sclance for Life. 2/6 amplify it with PCR (© 2007 Pearson Prentice Hall Inc « We then treat the DNA with restriction enzymes - cut DNA at specific sequences- Everyone's DNA is different, so everyone’s DNA will cut at different sites * This results in different sized fragments * The different sized fragments are called restriction fragment length polymorphisms, or RFLPs Variable * We can observe the mw = BRE wer different sized fragments in eee: peer an experiment that . Carer] separates DNA based on —~Smrmmm-l2 AVNTRs fragment size called Gel Electrophoresis ~Sem sures Student 1 * Everyone has genetic sequences called variable ~Smmmmme{-— cvntrs number tandem repeats, or VNTRs heme | sums " Student 2 ~ Everyone has different Homologous chromosomes amounts of VNTRs egret sma - The VNTRs make the different sized RFLPs Gel Electrophoresis 6 * Fragments of DNA from restriction enzyme cleavage are separated from each other when they migrate through a support called an agarose gel - It is similar to the yummy food Jell-O gelatin- It is actually made out of some of the same ingredients Cut DNA from Cut DNA from Power Student! Student2 DNA from two different individuals is cut with supply Large restriction enzymes and loaded on an agarose - i i fragments gel.When these fragments are subjected to an wins |7 0fDNA electric current, shorter fragments migrate ~~ RARA 6 through the gel faster than do larger fragments. 5 wew ih 4 Student 1 has DNA sequences that carry 4 and 5 u 3 repeat sequences. Student 2 has 3 and 6 repeats. * . The remaining DNA is DNA that does not carry repeat sequences. Even though the DNA is visible garere ga Small ge inthis figure, DNA is not visible with the unaided . ofpna = “Y" The size-based Ctrpeeuonreetemke separation of Molecules of DNA separate based on size when an electric current is applied to an agarose gel. * This method is called gel electrophoresis Gel Electrophoresis * The separated DNA fragments are then drawn out of the gel using a nylon membrane* The nylon membrane is treated with chemicals that break the hydrogen bonds in DNA and separate the strands Cut DNA from Cut DNA from Student 1 Student 2 Radio- RIFF F |7 active ‘ probe SNwEua The DNA is transferred from the gel onto filter paper and chemically treated to make it single stranded. The DNA is not visible on the filter paper and must be probed with single-stranded, radio- actively labeled DNA that is complementary to the repeat sequence. DNA that does not contain the repeat sequence will not bind to the probe. *-* The single stranded DNA Filter paper jc cross linked to the nylon membrane. 2007 Pearson Pence Malkin - By heat or UV light * Incubate the nylon membrane with a radioactive probe of single stranded DNA complementary to the VNTRs.Southern Blotting © crite ona ‘Cut with restriction enzyme @ Restriction tragments of longtna deteroed by Ieetton of recouraten - sequences Ierrecmtevon af name ~~ Agro | © soteiectrophoresis f —_ EZ. ay oo After staining with ethicium Sromide, DMA Iengmenta are visibie with UV tlkeninatioa © Tenster 10 Weight mee ier Soutnam blot Membrane ‘woneiaue r Get Se + Bioteng pape | SS Trey containing: ee butler olution © OKA ragmanis ransterres ‘exactly they were ranged in ngaroae gel Hybrickze with labeled probe © ona raamons compemantary tame prebe are viele liner nucaradiogtaony 0° ‘hemtuminesceoce A Southern blot is a method used in molecular biology for detection of a specific DNA sequence in DNA samples. Southern blotting combines transfer of electrophoresis-separated DNA fragments to a filter membrane — and subsequent fragment detection by probe hybridization.Famous cases solved by DNA fingerprinting DNA Fingerprints, stray cigarette helped solve 1997 killing, sheriff Says Amber Gail Creek, 14, was found dead in a Burlington marsh Racine County Sheriff Christopher Schmating on Tuesday Identified James P. Eaton as the 36-year-old Illinols man authorities suspect killed Amber Gall Creek, a 14-year-old runaway girl from Illinois, in 1997. Credit: Racine County Sheriff's Department The 1997 killing of a 14-year-old girl whose body was found in a Racine County marsh has been solved 17 years later with old fingerprints and new DNA evidence unknowingly provided by the allegedChristopher Schmaling identified the suspect as James P.Eaton, a 36-year-old Ilinols man, and sald he Is being held on $1 million ball in the death of Amber Gail Creek, an Illinois runaway. “Over the last 17 years we have dedicated thousands of investigative hours to bring this tragic and senseless murder to its resolution,” Schmaling told reporters at a news conference. "Today that day Is here." The break in the case stemmed from fingerprints left on the black Dlastic hag used to suffocate the teen, Schmaling sald, and Investigators were able to track Eaton and pull DNA evidence off one of his cigarettes. Eaton, a Palatine, Ill, resident, faces charges in Racine County of first-degree intentional homicide and hiding a corpse. Schmaling, who did not take questions from reporters, sald Amber's family has asked for privacy. Amber ran away from a state- operated juvenile shelter in Chicago on Jan. 23, 1997. She attended a party ata motel in Rolling Meadows, Ill, the week of her death and was last seen leaving the party and getting into a luxury car with a man. On Feb. 9, 1997, two men stumbled upon her body while they were searching for hunting sites at the Karcher Wildlife Refuge inthe Town of Burlington. She had been sexually assaulted, and her body was found posed with an upraised hand and the greeting "HI" written on the back of her hand, according to Journal Sentinel archives. She was found witha $5 Drice tag on her arm from a bookstore In the Schaumburg, lll, area. Investigators spent more than a year and a half examining more than 1,300 missing persons files before they Identifiedher, and the case drew national attention at the time, including a brofile on "America's Most Wanted" that aired Dec. 12, 1998. Fingerprints taken from the plastic bag and DNA evidence from Amber's body initially yielded no matches, despite the help of the FBI and agencies around the country. Then this year, through new fingerprint Identification technology used bya crime lab in Oktahoma, authorities were able to match the prints found on the plastic bag to Eaton's prints, Schmaling sald. Eaton had been fingerprinted in Illinois for past, minor offenses, but his name Dreviously had not come up in the Amber Creek case. After learning of the fingerprint match on Feb. 28, Racine County Investigators conducted surveillance on Eaton for several days, and when he dropped a cigarette while waiting for a train in the Chicago area, they picked it up for DNA evidence. Itmatched the DNA found on Amber's body, Schmaling sald."Our sense of accomplishment is tempered by the pain and loss we know Amber's family continues to confront every day,” Schmaling said. "Our thoughts and prayers remain with Amber's father, mother, other family and loved ones." Itwasn't clear whether Amber and Eaton knew each other or how they first would have come into contact. Eaton was 19 at the time of the killing. The sherlif sald Investigators are still looking for people who can Drovide with Information in the case. They are encouraged to contact the Racine County sheriff's office at (262) 636-3225 or leave an anonymous tip at Crime Stoppers of Racine at (888) 636-9330.References Wwww.bcps.org http://www. n. https://en.wiki ja.orOs ae : si i i i= Terms related to DNA fingerprinting ; Reference samplePolymorphismJunk DNA ET 95%gene Tandem repeat 10 = 14 - 10Restriction endonucleasencinnite Collection & preservation of biological samplesAgere r RFLP = Ss fo] — ->—> DNA Restriction \. <_ ™ Electrophoresis extraction digestion ~ Transfer of DNAto membrane — meray Hybridization of DNASouthern blotting Ty pes « f DNA fingerprinting RPLPDIY ©) ol ee 4D | mec’ €2| lilies $s" EX Ill 1a