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How they make a scrimillion copies of the DNA Amplification Capillary Electrophereses Interpretation.

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Presentation on theme: "How they make a scrimillion copies of the DNA Amplification Capillary Electrophereses Interpretation."— Presentation transcript:

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2 How they make a scrimillion copies of the DNA

3 Amplification Capillary Electrophereses Interpretation

4 Find the areas of interest D8S1179 D21S11 Amelogenin D18S51 D3S1358 FGA vWA D5S818 D13S317 D7S820

5 D8S1179 D21S11 Amelogenin D18S51 D3S1358 FGA vWA D5S818 D13S317 D7S820 Find the areas before and after the areas of interest (primer binding sites)

6 So weve removed the DNA from the cells, now what? PCR me, ASAP!

7 P C R PCR = Polymerase chain reaction. Used to make scrimillions of copies of the loci of interest.

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13 bases = nucleotides

14 CGACATGTCAT TACGTAGGCTA ACTGCTACAGT GATAGCAGCGT CCGATCTAGCA

15 G T C T C A G A

16 G T C T C A G A

17 G T C T C A G A

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19 G T C T A G A G T C T C A G A G C T C T C A G A

20 G T C T A G A G T C T C A G A G C T C T C A G A

21 G T C T A G A G T C T C A G A G C T C T C A G A

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23 So we can just keep splitting it open and adding bases and make millions and millions of copies. This is amplification. Technically, this is not a science term either. Amplify: Root words Amplus, large + facere, to make* *Websters New World Compact School and Office Dictionary, at 15, (1989)

24 Holt Science and Technology, Life Science, p. 137 The loci we care about are tagged with a flourescent dye...

25 Getting to the Test Result

26 What do you do with a billion copies of markers? You inject them into the 310 and get a reading.

27 ABI 310 Genetic Analyzer: Capillary Electrophoresis Slide courtesy of Simon Ford Detector Window Slide courtesy of Simon Ford

28 Okay, now what? Meet the ABI Prism 310 Genetic Analyzer Slide courtesy of John Butler

29 Put on your thinking caps and open your eyes...

30 What we care about: When did it jump, and how high? Length aka number of repeats? Quantity aka how much DNA do they have?

31 What we care about: When did it jump, and how high? Length aka number of repeats...

32 Length The smaller DNA pieces pass through the straw (aka capillary) faster than the bigger pieces. They are recorded first. Slide courtesy of Carll Ladd Picture courtesy of Carll Ladd

33 Why do we care about this? Because they go through the capillary according to size, we know what comes through when. So we can label them.

34 AmpFlSTR ® Identifiler (Applied Biosystems)

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38 AMEL D3S1358 TH01 TPOX D2S1338 D19S433 FGA D21S11 D18S51 CSF1PO D16S539 D7S820 D13S31 7 D5S818 VWA D8S1179 See the funny looking ruler at the top? That measures the number of base pairs. X is about 106 base pairs long. The 24 at D2 is 344 base pairs long. AmpFlSTR ® Identifiler (Applied Biosystems) Slide courtesy of John Butler Why do we care about how long these things are? The length is one of the things that tells us what these things are.

39 AMEL D3S1358 TH01 TPOX D2S1338 D19S433 FGA D21S11 D18S51 CSF1PO D16S539 D7S820 D13S31 7 D5S818 VWA D8S1179 The importance of color... AmpFlSTR ® Identifiler (Applied Biosystems) Slide courtesy of John Butler

40 Quantity of DNA The amount of DNA is measured on the right.

41 WHAT DOES THIS MEAN?? The DNA profile found at the crime scene Matches the DNA profile said to be from the defendant...

42 Lets compare DNA profiles: Stand up! (Humor me, would ya?) Match is with female With white skin And blue eyes And red hair Who was raised catholic By an Irish mother And has one child

43 Whos in the mix? Its time to stand up!! Match is with female or male With white skin or olive skin And blue eyes or brown eyes And red hair or black hair Who was raised catholic or free spirit By an Irish mother or an Italian one And has one daughter or one son

44 Whos better than you?

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46 State of Texas v. Josiah Sutton (1999) Woman raped in car by two men Identifies Sutton and Adams DNA test on vaginal swab, pubic combings, jeans, and semen stain on car seat

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48 Trial testimony Everyone has a unique DNA pattern Suttons pattern was found in the vaginal swab, pubic combings, and jeans No testimony about semen on car seat (#1 unknown sample)

49 Whos better than you? Its time to Do!

50 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13

51 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defend ant 12,13 Alleles Present 10,12,13,14, 15

52 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

53 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

54 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

55 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

56 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

57 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

58 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

59 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

60 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

61 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

62 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

63 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

64 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

65 8,8 8,9 8,10 8,11 8,12 8,13 8,14 8,15 8,16 8,17 8,18 8,19 9,9 9,10 9,11 9,12 9,13 9,14 9,15 9,16 9,17 9,18 9,19 10,10 10,11 10,12 10,13 10,14 10,15 10,16 10,17 10,18 10,19 11,11 11,12 11,13 11,14 11,15 11,16 11,17 11,18 11,19 12,12 12,13 12,14 12,15 12,16 12,17 12,18 12,19 13,13 13,14 13,15 13,16 13,17 13,18 13,19 14,14 14,15 14,16 14,17 14,18 14,19 15,15 15,16 15,17 15,18 15,19 16,16 16,17 16,18 16,19 17,17 17,18 17,19 18,18 18,1919,19 Defendant 12,13 Alleles Present 10,12,13,14, 15

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