1. Lab 18 DNA Murder Mystery
BY-102
Spring 2013
Zach Nolen

2. DNA Fingerprinting
Is a technique used by forensic scientists to identify people based on their DNA profiles
The key to DNA fingerprinting is Restriction Fragment Length Polymorphisms (RFLPs)

3. When is it used? Crime Investigations Familial Relationships Rape
Violent Crimes
Familial Relationships
Paternity tests

4. How Specific Are RFLPs?
Under normal circumstances, no other human has your exact complement of genes, unless…
You are an identical twin

5. Murder Case

6. Murder Case
After going missing, Lynda Mann, a 15-year old schoolgirl was raped and murdered in the grounds of Carlton Hayes psychiatric hospital in Narborough, Leicestershire, in November Forensic examination of semen sample that it was a type found only in 10% of men, and was from someone with type A blood. However, the police did not find a suspect. In 1986, another 150year old schoolgirl, Dawn Ashworth, was similarly sexually assaulted and strangled in the nearby village of Enderby, and semen samples showed the same blood type.

7. Suspect 1 Richard Buckland (17 years old)
Local with learning disabilities
Worked at Carlton Hayes psychiatric hospital
Seen near murder scene of Dawn Ashworth
Knew detailed information regarding body of deceased
Confessed to the murder of Dawn Ashworth but not to the murder of Lynda Mann

8. Suspect 2 Colin Pitchfork (27 years old) Local baker
Previous convictions for flashing
Paid someone to give a false semen sample to police during the investigation supposedly because of his prior convections.

9. How can we find the culprit?
DNA Fingerprinting

10. Steps Extract DNA Amplify DNA Cut DNA Sort DNA Denature DNA
Hybridize DNA
Make an X-Ray

11. Extract DNA
We performed an extraction last week using cold ethanol. Keep in mind that there are other ways to extract DNA.
We will be going through these steps using the DNA sheet that you should have printed out.
If you do not have the sheet, I have a few extras.

12. Amplify DNA We amplify DNA using Polymerase Chain Reaction (PCR).
This allows us to increase the total amount of DNA that we have
For our exercise, everyone having their copy of DNA will act as the amplification

13. Cut DNA NeoSci 1 5’ GG|CC 3’ 3’ CC|GG 5’ NeoSci 2 5’ GA|AG 3’
To cut DNA, Restriction enzymes are used.
These enzymes cut DNA by recognizing specific sites in chains of DNA
We will act as restriction enzymes and cut our DNA at the indicated sites.
Make sure that the sites match exactly
NeoSci 1
5’ GG|CC 3’
3’ CC|GG 5’
NeoSci 2
5’ GA|AG 3’
3’ CT|TC 5’

14. Sort DNA
After the DNA is cut using restriction enzymes, it must be sorted using gel electrophoresis
This process separates DNA fragments based on size
We will cut paper DNA fragments out ant place them on our paper gel
Fill out p. 215 by placing dark lines at any location where a fragment would be present

15. Denature DNA
After the DNA has been sorted, it must then be denatured using either heat or chemical.
To denature our DNA, we will remove the yellow strips from the bottom of the fragments in our lanes

16. Hybridize DNA
This step binds a radioactive probe to the fragmented DNA so that it can be seen.
We will cut out the “T C C” probes and place them under fragments with the corresponding letters “A G G”

17. Make an X-Ray
The process of taking an X-Ray of hybridized DNA is called Autoradiography
Fill in lines on p. 214 to indicate fragments where hybridization has occurred.

18. Results Suspect 2: Colin Pitchfork is the murder.
Colin Pitchfork was the first person ever to convicted of murder based on DNA fingerprinting evidence.

19. Paternity Tests

20. Before you leave lab
Clean up your work station

21. Before next lab Read Lab 7 – The Cardiovascular System
Do Online Prelab
Complete sections in lab manual for lab notebook check