Daniel Hering RFLP Electrophoresis (DNA Fingerprinting)

Quantitative Analysis of DNA Fragment Sizes Quantitative Analysis of DNA Fragment Sizes
Lambda/HindIII size marker Band 1 2 3 4 5 6
Actual size (bp) Distance (mm) 5 4.5 4 3.5 3 2.5 2 1.5 1 0.5 0 10 15 20

Distance (mm)

y = -0.049x + 5.1033 Series1 Linear (Series1)

23,130 9,416 6,557 4,361 2,322 2,027

18.3 21.2 24.5 28.7 36.1 37.5

25

30

35

40

LOG Apprix. Size (bp)

Crime Scene
Distance (mm) Approx. size (bp)

Suspect 1
Distance (mm) Approx. size (bp)

Suspect 2
Distance (mm) Approx. size (bp)

Suspect 3
Distance (mm) Approx. size (bp)

30 32.8 47.8

4298 3134 577

33.9 45.1 48

2768 782 564

33.1 39.5 44.7

3030 1472 818

30 32.8 47.8

4298 3134 577

Suspect 4
Distance (mm) Approx. size (bp)

Suspect 5
Distance (mm) Approx. size (bp)

32.9 45.5 56.6

3099 748 214

34 37.8 46

2737 1783 707 204

57

Reflection Questions RFLP Electrophoresis (DNA Fingerprinting)

1. Refer to your results table. Which of the DNA samples have the same number of restriction sites? Samples 1-4

2. Which sample has the smallest DNA fragment? Which has the largest? Smallest DNA fragment: Sample 5 Largest DNA fragment: Sample 3

3. Compare the fragment sizes of the suspects and the crime scene. Which samples match the crime scene DNA? Which samples do not match the crime scene DNA? Sample 3 matches the crime scene Samples 1-2 & 4-5 do not match the crime scene

4. What does it mean if a suspect matches the crime scene? What does it mean if a suspect does not match the crime scene? The suspect matches the crime scene when the DNA of the suspect and the DNA found at the crime scene exhibit similar placement of fragmentation, meaning two samples contain the same DNA. The suspect does not match the crime scene when the fragments are placed in different spot than the crime scene DNA fragments.

5. What would your gel look like if the DNA were not fragmented? You would have a continuous line of DNA running along the gel.

6. What determines where a restriction endonuclease will cut a DNA molecule? The specific code sequence of the DNA that the restriction endonuclease is looking for (life.illinois.edu).

7. Electrophoresis is one method of separating molecules. Paper chromatography is another method of separating molecules. What are two similarities and two differences between these two methods? Electrophoresis and chromatography carry out the process of extraction to compare a certain two different things. However, electrophoresis is used to compare DNA samples, while chromatography is used to compare different types of plants.

8. If you were called away during the electrophoresis procedure and were not able to monitor your electrophoresis run, what do you think would happen if the electricity were to remain running in your absence? The DNA samples would keep forming fragments and there would be fragments all over the gel, making it difficult to compare any samples.