Science | Laboratory Instructions | DNA Laboratory Instructions Laboratory: DNA Materials Supplied Student Guide Laboratory Guidelines Electropherograms (online) Codis STR Table (online and print) Also Needed scientific calculator (optional) Safety Review the Laboratory Guidelines before conducting the lab. Crime Summary On a Midwest college campus, three female students were assaulted within a six-month period. Each woman was beaten, robbed, and left for dead. All three women survived the attacks. During the last attack, the perpetrator dropped a bloodstained knife, which was recovered by police. When questioned by police, each woman said that the attacker was a tall man who wore gloves and a mask. He threatened them with a knife and took them off the main campus quad into a wooded area where he beat them. Although they could not see the attacker’s face, each woman did manage to scratch him. Police investigators took scrapings from under their fingernails, blood samples, and their bloodstained clothes for DNA analysis. In the course of the investigation, police detained four possible suspects. All the suspects are white males. Although Suspects A, B, and C could not be held because the victims could not identify them from a lineup, police did manage to get saliva samples. Unbeknownst to the suspects, police collected discarded cigarette butts and drinking glasses so the saliva could be used for DNA testing. Suspect D was arrested shortly after the last assault. He had scratch marks on his arms, and police confiscated a bloody pair of jeans from his apartment. They took DNA samples from him and the clothing. They have sent the evidence to you, the forensic scientist, to conduct the analysis. © 2012 K12 Inc. All rights reserved. Copying or distributing without K12’s written consent is prohibited. Page 1 of 4 Science | Laboratory Instructions | DNA Part 1: Use an electropherogram to create a DNA profile. You have run the PCR with STRs for all of the evidence samples and suspects. You have used the following STRs: Amelogenin D3S1358 vWA FGA D18S51 You have separated the PCR product DNA fragments by capillary electrophoresis and have obtained the electropherograms corresponding to each sample. Go online to locate the electropherograms. They are organized across five screens into the following sets: DNA from Victims DNA from Victims’ Shirts DNA from Victims’ Fingernail Scrapings DNA from Knife and Jeans DNA from Suspects Study the electropherogram for Victim 1. The first thing you will notice is that the number of STRs (allele repeat number) at each marker is not available. Only the base pair length is available. Computers generally calculate the number of STRs from the base pair length, but in this lab, you will do the work of the computer. Let’s walk through the process. 1. For each STR marker, locate the base pair length (bp on the horizontal axis) for the peak or peaks. For example, for Victim 1, there is one peak for the STR marker D18S51. The peak is located at bp 272. 2. Now use the CODIS STR Marker table. Click Codis STR Table in the upper right-hand corner of the electropherogram to access the table online. You can also use the print version of the table. To convert the bp number to the allele repeat number: Locate the base pair length in the appropriate STR marker column. Read the allele repeat number in the same row. For example, the D18S51 peak for victim 1 has a bp value of 272. Find the row with 272 under the D18S51 column. Then read the value in the same row under the Allele (repeat #) column, which in this case is 9. Example: Allele repeat number for Victim 1, D18S51 Base pair length (bp) Allele (repeat #) Amel D3S1358 7 8 99 103 D18S51 264 268 9 107 272 © 2012 K12 Inc. All rights reserved. Copying or distributing without K12’s written consent is prohibited. vWA FGA Page 2 of 4 Science | Laboratory Instructions | DNA Note that Victim 1 is homozygous for D18S51 (one peak). That is, if you find one peak for STR (D18S51), then you know she inherited STR D18S51 (with an allele repeat number of 9) from BOTH parents. This is recorded as (9, 9) in the DNA profile. If there are two peaks nearby each other for STR, then you know she is heterozygous for this STR, and you need to locate the Allele repeat number for each peak. Notice below that FGA, for example, has two Allele repeat numbers, one for each peak on the electropherogram. Your job now is to complete the DNA Profile Results: Table of Alleles below. The DNA profile of Victim 1 for all five STR markers has been recorded for you. Study the values and the electropherogram for Victim 1 to make sure you understand the process. When you have finished the table, go to the next part of the lab. DNA Profile Results: Table of Alleles Sample Base pair length (bp) Amel (XX or XY) D3S1358 vWA FGA D18S31 XX 11, 12 15, 15 13, 14 9, 9 Victim 1 Victim 2 Victim 3 Victim 1 shirt Victim 2 shirt Victim 3 shirt Knife Suspect D jeans Victim 1 fingernail scrapings Victim 2 fingernail scrapings Victim 3 fingernail scrapings Suspect A Suspect B Suspect C Suspect D Part 2: Draw conclusions. Examine the DNA profile results table. 1. Which suspects can you exclude? 2. Which suspects can you include? 3. Can you associate any of the suspects with specific victims? If so, which combinations? © 2012 K12 Inc. All rights reserved. Copying or distributing without K12’s written consent is prohibited. Page 3 of 4 Science | Laboratory Instructions | DNA 4. Are there any inconsistencies? Part 3: Search CODIS database. Compare the DNA profiles from this case to those in the CODIS database below. Sample codes that start with E stand for evidence, while those that start with O stand for offenders or suspects who were arrested. Do any of the DNA profiles from your case match any in the CODIS database? If so, which match occurs, and what is the CODIS case? CODIS Database Sample Amel E00100009 E00106010 E00260050 E03010025 E00562110 O00106530 O00352130 O01106220 O01897830 O05534297 Sample E00100009 E00106010 E00260050 E03010025 Vaginal swab, unsolve E00562110 O00106530 O00352130 O01106220 O01897830 O05534297 D3S1358 XY XX XY XY XY XX XY XY XY XY 16, 17 12, 13 14, 14 14, 14 19, 20 18, 18 15, 15 17, 17 13, 14 20, 20 vWA 14, 15 19, 19 18, 19 16, 16 13, 14 14, 15 12, 12 15, 15 20, 20 18, 19 FGA 25, 25 13, 14 24, 24 20, 21 22, 23 19, 19 17, 18 24, 25 22, 23 20, 21 D18S51 10, 11 12, 12 7, 7 10, 10 15, 15 11, 11 16, 16 15, 16 12, 13 7, 8 Description Bloodstain, unsolved murder in Pittsburgh, PA Bloodstain, unsolved murder in Washington, DC Oral swab, unsolved sexual assault in Tampa, FL Vaginal swab, unsolved rape case in Yuma, AZ Vaginal swab, unsolved rape case in Los Angeles, CA Sexual offender in Chicago, IL Robbery suspect in Raleigh, NC Burglary suspect in Houston, TX Sexual offender in New Orleans, LA Vehicular manslaughter suspect in Seattle, WA © 2012 K12 Inc. All rights reserved. Copying or distributing without K12’s written consent is prohibited. Page 4 of 4