of 65 /65
How they make How they make a scrimillion a scrimillion copies of the copies of the DNA DNA

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

Embed Size (px)

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

  • Slide 1

Slide 2 How they make a scrimillion copies of the DNA Slide 3 Amplification Capillary Electrophereses Interpretation Slide 4 Find the areas of interest D8S1179 D21S11 Amelogenin D18S51 D3S1358 FGA vWA D5S818 D13S317 D7S820 Slide 5 D8S1179 D21S11 Amelogenin D18S51 D3S1358 FGA vWA D5S818 D13S317 D7S820 Find the areas before and after the areas of interest (primer binding sites) Slide 6 So weve removed the DNA from the cells, now what? PCR me, ASAP! Slide 7 P C R PCR = Polymerase chain reaction. Used to make scrimillions of copies of the loci of interest. Slide 8 Slide 9 Slide 10 Slide 11 Slide 12 Slide 13 bases = nucleotides Slide 14 CGACATGTCAT TACGTAGGCTA ACTGCTACAGT GATAGCAGCGT CCGATCTAGCA Slide 15 G T C T C A G A Slide 16 G T C T C A G A Slide 17 G T C T C A G A Slide 18 Slide 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 Slide 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 Slide 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 Slide 22 Slide 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) Slide 24 Holt Science and Technology, Life Science, p. 137 The loci we care about are tagged with a flourescent dye... Slide 25 Getting to the Test Result Slide 26 What do you do with a billion copies of markers? You inject them into the 310 and get a reading. Slide 27 ABI 310 Genetic Analyzer: Capillary Electrophoresis Slide courtesy of Simon Ford Detector Window Slide courtesy of Simon Ford Slide 28 Okay, now what? Meet the 310... ABI Prism 310 Genetic Analyzer Slide courtesy of John Butler Slide 29 Put on your thinking caps and open your eyes... Slide 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? Slide 31 What we care about: When did it jump, and how high? Length aka number of repeats... Slide 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 Slide 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. Slide 34 AmpFlSTR Identifiler (Applied Biosystems) Slide 35 Slide 36 Slide 37 Slide 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. Slide 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 Slide 40 Quantity of DNA The amount of DNA is measured on the right. Slide 41 WHAT DOES THIS MEAN?? The DNA profile found at the crime scene Matches the DNA profile said to be from the defendant... Slide 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 Slide 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 Slide 44 Whos better than you? Slide 45 Slide 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 Slide 47 Slide 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) Slide 49 Whos better than you? Its time to Do! Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 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 Slide 66