3-17-10 Biotechnology-1-color - Clicker Question _____ is...

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Unformatted text preview: Clicker Question _____ is the process in which RNA is synthesized and _____ is the process in which protein is synthesized synthesized. A) B) C) D) E) Where are we? Last Last Time I Talked about… DNA, DNA, RNA, proteins, replication, transcription, translation and protein chemistry. Today and next time ti I will discuss biotechnology will and how we can use our knowledge to cross the genetic frontier to manipulate genes and produce proteins for our benefit. I will also discuss the PCR will technique that you will use in lab. Translation, transcription Translation, transfection Transcription, translation Transliteration, translation Transnucleation, transformation The Genetic Frontier While While people long for the security of security of civilization, we are also curious about curious about what is yet to be known at its frontiers. At At the frontiers, which are always changing, there can be hope and hope there can be horror. horror Knowledge Knowledge unveiled or revealed at the genetic frontier will change what it means to be human. Aldous Huxley, Author of Brave New World, is T. H. Huxley’s Grandson and Andrew Fielding Huxley’s Half Brother Where do we come from? What are we? Where are we going? Right and Wrong “The student in high school, in college and in graduate school must be concerned, in part at least, with the words ‘right’ and ‘wrong’ in both the ethical and the mathematical sense.” --James Bryant Conant in General Education in a Free Society General Education in a Free Society time.com 1945 time.com 1 An Ethical Covenant? Marshall Nirenberg, Who Discovered the Relationship Between Codons and Amino Acids, Wrote… Howard Gardner, who developed the idea of multiple intelligences, proposes that scientists enter an ethical ethical covenant stating they have a prepared mind to focus on the possible applications or misapplications of their work. http://nobelprize.org/nobel_prizes/medicine/laureates/1968/nirenberg-interview.html http://www.howardgardner.com/Papers/documents/Eth%20Resp%20of%20Sci_Feb-02_HG%20pdf.pdf Will Society be Prepared? Homo Sapiens—Wise Humans Homo Sapiens—Wise Humans www.bioethics.gov www.ucsusa.org Life Learning Devices Centuries of Wisdom—at a Touch of a Button lifelearningdevices.com www.whitehouse.gov www.nytimes.com 2 Science: Wisdom? Politics? On On February, 2004, Elizabeth Blackburn was dismissed by President Bush from the President’s Council on Bioethics, perhaps due to her Bioethics, perhaps due to her dissenting dissenting view on stem cells. In In June, 2009, President Barack Obama disbanded the President’s Council on Bioethics because it was “a philosophically leaning advisory group.” Science: Science: Wisdom? Politics? On On November 24, 2009, President Obama created a new Presidential Commission for the Study of Bioethical Issues that “offers practical policy options”. li President President Obama said, “As our nation invests in science and innovation and pursues advances in biomedical research and health care, it’s imperative that we do so in a responsible manner.” …but is there a shared trust? James Watson Speaks His Mind "It's my impression that none of the genomeproject leaders have gotten up and said, 'What we are going to do with this information; I think we should use think we should use it’. Maybe they're afraid of offending people." "Going for perfection was something I always thought you should do. You always want the perfect girl." "Who wants an ugly baby?" “So, “So, just to be equally provocative and to even the score, so to speak, let me juxtapose to Watson’s list a few other trait modifications that some people may want to carry out…. Breeding young males as killing machines Breeding young males as killing machines…. Turning Turning normal people into obedient servants…. Creating Creating young women who will function as docile sex slaves; Breeding subBreeding a sub-class of manual laborers with distinctive physical traits…. Then what?” “Our “Our technological capacity to genetically-engineer human and geneticallyanimal traits is developing rapidly. Do we have the institutional capacities in place to manage the associated risks?” www.leiss.ca Anna Kournikova Toronto Globe and Mail 26 October 2002. 3 The Future is Here: Dr. Jeffrey Steinberg of The Fertility Institutes www.fertility-docs.com Designer Babies http://www.fertility-docs.com/news_events.phtml?ID=22 Interview with Dr. Jeffrey Steinberg January 29, 2009 'pity this busy monster, manunkind' The Birthmark by by Nathaniel Hawthorne “A “A world of made is not a world of born” world of born -e. e. cummings A story about a scientist who tries to make his wife perfect. www.online-literature.com 4 How Did We Get Here? Majority View of Evolution: A Gradual Vertical Evolution: Gene Transfer with Near-Perfect Fidelity NearBio G 1110 Students Minority View: Richard B. Goldschmidt— Goldschmidt— Macroevolution Does Not Result from Gradual Changes but from Drastic Changes Perhaps Richard Goldschmidt, One Person Who was Not Appreciated in his Lifetime, was Right Evolution Via Horizontal Gene Transfer When When I talked about reproductive biology, I talked about vertical vertical gene transfer from parent to offspring, and I assume that vertical gene transfer with near perfect fidelity along with natural selection gave rise to the various selection gave rise to the various extinct extinct and extant taxa in the living world. Today Today I will present historical evidence suggesting the possibility of horizontal gene horizontal transfer, and the methods used methods to transfer genes with desired traits from any organism or taxon into another. What are Viruses? www.nsf.gov I mentioned in the first lecture that the botanist Martinus Beijerinck (1898) thought viruses found in a cellcell-free filtrate were the smallest form of life and that the chemist Wendell Stanley (1935) thought they were the largest chemicals. 5 Cancers May Be Transmitted from Organism to Organism by Viruses Peyton Peyton Rous (1911) described a cancer in chickens that could be transmitted from be transmitted from chicken chicken to chicken. He He found that “small “small quantities of a cellcellfree filtrate have sufficed to transmit the growth to susceptible fowl.” Genes Can Move Horizontally From One Taxon to Another Hermann Hermann Muller (1922) postulated that viruses may be genes. Benjamin Benjamin Duggar and Joanne Armstrong (1923) suggested that viruses are “rebellious genes that have “rebellious escaped from the chains of escaped from the chains of coordination.” coordination.” E. E. B. Wilson (1925) postulated that the genetic system responsible for the development and evolution of organisms could be affected by viruses. Indeed, Indeed, viruses can cause “horizontal” “horizontal” gene transfer between organisms and taxa making us all part of a genetic web of life. The Gene for Virulence Can Be Transferred From a Dead Organism to a Live One in a Process Known as Transformation Fred Griffith (1928) demonstrated the process of horizontal gene transfer between organisms of the same species when he showed that transformation transformation occurred between between dead smooth virulent and live rough avirulent Pneumococci. Pneumococci. Engineered Horizontal Gene Transfer Stanley Stanley Cohen and Herbert Boyer developed and patented a technique to introduce horizontal gene transfer. Use Use either a plasmid or a plasmid deactivated deactivated virus to facilitate the transfer. A plasmid is a small circular DNA plasmid molecule found in a bacterium that replicates faster than its large DNA molecule or “chromosome”. Steps in Genetic Engineering Developed and Patented by Herbert Boyer and Stanley Cohen Choose and isolate the Choose and isolate the gene of interest from an organism that has the desired trait. Insert Insert the gene into (transform) bacteria that multiply and divide rapidly so that the gene gets cloned gets cloned. Isolate Isolate the protein of interest from the reproducing bacteria; or… Isolate Isolate the gene of interest from the reproducing bacteria and use it to transform transform eukaryotic organisms, including cultured cells, plants, mice, sheep, cows and humans. Cloning a Gene: Making Recombinant DNA Isolate plasmid Isolate a plasmid. Insert Insert the gene of interest into the plasmid. Insert Insert a gene for antibiotic antibiotic resistance resistance as a selectable marker along with the gene of interest. Recombinant Recombinant DNA is DNA is that has been produced from the combination of two or more DNA sources. 6 Cloning a Gene: Making Many Copies of a Gene Transform Transform bacterial cells with the recombinant DNA. DNA. Grow Grow the bacteria in the presence of the antibiotic that antibiotic will kill the untransformed cells will kill the untransformed cells, but not the transformed cells. The The transformed cells multiply multiply and divide forming many copies (clones) of the gene. The The bacteria then serve as an abundant abundant source of the cloned gene protein gene or protein encoded by the cloned gene. Werner Arber, Daniel Nathans and Hamilton Smith Discovered that you Can Cut the Gene of Interest Out of the Genome with Restriction Enzymes Restriction Restriction Enzymes: A Bacterial Defense System Bacteria Bacteria produce enzymes, known as restriction restriction enzymes that naturally protect the bacterial cells from attacks by viruses, known as bacteriophages or phages. phages The enzymes restrict the growth of the phages. restrict The restriction The restriction enzymes are named after the bacterium from which they were isolated (e.g. Eco R1 comes from th R1 E. E. coli—a bacterial species found in our colon). The restriction The restriction enzymes recognize a sequence of 4-8 4nucleotides in the phage’s DNA and cuts it, inactivating the phage and restricting its ability to infect the restricting bacterium. The The bacterial DNA sequences that would be cut by the restriction enzyme are methylated so that the enzyme does not recognize and cut the host sequence. Restriction Enzymes Can Be Used as Molecular Scissors The The restriction enzymes recognize a sequence of 48 nucleotides in any DNA and can be used to cut it at this site. The The restriction enzymes cut the sequence so as to leave two “sticky ends”. Restriction Restriction enzymes are enzymes that act as molecular molecular scissors once they come in contact with the correct substrate and bind with high affinity. 7 DNA Fragments of Different Sizes Can Be Separated by Electrophoresis Separation of DNA By Electrophoresis After Digestion with a Restriction Enzyme (SmaI) The The restriction enzymes cut DNA from a given organism into manageable pieces. The The DNA is negatively charged and the various-sized variouspieces can be separated by gel electrophoresis. gel Creating Recombinant DNA with a Restriction Enzyme and DNA Ligase A DNA fragment containing the gene of interest DNA along with nearby genes are isolated from their native DNA using a certain restriction enzyme. restriction The plasmid is cut with the same restriction The plasmid is cut with the same restriction enzyme enzyme to make complementary “sticky complementary ends”. When When the DNA fragment containing the gene of interest is mixed with the cut-open plasmid, the cutbases at the cut ends of the two complementary ends of the separate DNAs pair. pair DNA DNA ligase is then used to covalently join the two DNA pieces. DNA Ligase Joins the Gene to the Plasmid to Form Form Recombinant DNA Transformation Bacteria Bacteria are then transformed transformed with the recombinant DNA plasmid. The transformed The transformed bacteria bacteria multiply and divide in the presence of an antibiotic to form a bacterial clone that carries many copies of the inserted gene. cDNA: Composed of Only Exons A gene isolated from gene eukaryotes may have introns introns that the prokaryotic bacteria are not able to splice out in the appropriate way. appropriate way. In In this case, one makes a gene with DNA that is complementary complementary to the fully spliced messenger RNA. This is called cDNA. cDNA Making Making DNA from RNA requires an enzyme known as reverse reverse transcriptase. 8 Reverse Transcriptase was Discovered in Retroviruses by David Baltimore and Howard Temin Reverse Transcriptase Retroviruses Retroviruses utilize RNA as RNA their genetic material. (HIV is a retrovirus). Inside Inside the host, the viral RNA is RNA transcribed into viral DNA with DNA reverse reverse transcriptase. The The viral DNA is DNA inserted into the DNA of DNA of the host. Use Use cDNA Approach to Eliminate the Introns from the Gene Isolate Isolate the messenger RNA of interest. messenger Add Add dATP, dGTP, dCTP and dTTP and the enzyme, reverse transcriptase, to make a single reverse DNA strand that is complementary to the mRNA. Eli Eliminate the RNA using RNase. th RNA RN Add DNA Add DNA polymerase and the deoxy nucleotides to synthesize the complementary strand of DNA. This results in double stranded cDNA that codes cDNA only for the expressed sequences or exons. ex exons The The cDNA is then inserted into a plasmid and the bacteria are transformed exactly as they are when genomic genomic DNA, isolated from the genome, is used. Transformation Vectors Bacteria Bacteria are typically transformed with plasmids or plasmids bacteriophage. Plants Plants are typically transformed with the Ti plasmid plasmid from Agrobacterium. Animals Animals are typically transformed with deactivated deactivated viruses. Genes Genes can also be shot into the cells with the gene gun, gene developed by John Sanford at Cornell and on display at the Smithsonian Institute. John Sanford: Gene Gun 9 Applications of Genetic Engineering Bacteria Bacteria or other cells containing the new gene can be cultured in a bioreactor so that they will bioreactor produce and secrete the protein coded by the gene of interest (e.g. insulin, EPO, human growth hormone, folliclestimulating hormone, interferon, cellulases used to process biofuels or the digestive enzymes used in laundry detergent). Many Many products are produced through the collaboration of biotech startups biotech startups and pharmaceutical companies. Herbert Boyer: Genentech (NYSE: DNA) Arthur Kornberg: DNAX Acquired by ScheringScheringPlough, which Merged in 2009 with Merck (MRK) On March 26, 2009, Roche acquired Genentech and DNA ceased being traded. http://www.gene.com/ www.roche.com http://summit.stanford.edu/newsevents/05_0415_kornberg.html www.merck.com Walter Gilbert: Biogen IDEC (NASDAQ: BIIB) “…that when we have the complete sequence of the human genome, we will know what it is to be human”. Genetic Engineering & Biotechnology News: GENdex http://www.biogenidec.com/ http://www.genengnews.com/ 10 Insulin Insulin injections are needed by some diabetics. Insulin Insulin used to be isolated from cows and pigs—it pigs— sometimes caused an allergic response in humans. Now Now recombinant “human insulin” is available and it does does not cause an allergic response. Human follicleHuman folliclestimulating hormone (FSH), used for infertility treatments used to be isolated from the urine of postpostmenopausal women. Post menopausal Post-menopausal women women have low levels of estrogen and progesterone and high levels of FSH and LH. Now “urineNow “urine-free” recombinant “human FSH” is available. lillydiabetes.com follistim.com Recombinant Erythropoietin Made in Erythropoietin Mammalian Cell Cultures Helps People with Kidney Disease procrit.com Human Human growth hormone (HGH) used to be isolated from human cadavers to treat very short people with a human growth hormone deficiency. Some of these people contracted Creutzfeldt-Jakob Creutzfeldtdisease, a prion disease. Now Now recombinant “human growth hormone” is available and there is no chance of contracting humatrope CreutzfeldtCreutzfeldt-Jakob disease. Recombinant Human Growth Hormone Helps Very Short People Grow Very Aging— Aging—A Human Growth Hormone Deficiency Disease? The The body’s production of human growth hormone declines with age. Can aging be defined in Can aging be defined in part part as a human growth hormone deficiency disease? If If so, should it be treated? Should health insurance cover www.s-hgh.com treatment? www.nutropin.com www.humangrowthhormone.me 11 Recombinant Vaccines can Prevent Hepatitis and other Diseases recombivax_hb Recombinant DNA Technology Makes Interferon Cheap and Accessible for the Treatment of Hepatitis and other Viral Infections PHRMA: The Value of Medicines “As a result of these new discoveries, medicines are taking on an increasingly important role in patient care. Because of this we are spending more on pharmaceuticals In more on pharmaceuticals. In return, return, more patients are living longer, better lives; overall health care costs are contained as patients avoid invasive surgeries and costly hospital and nursing home stays; and the economy is strengthened through improved worker productivity.” pharmamkting pegintron.com www.phrma.org What are the Long Term Goals of our Healthcare Policies? Are Are we defining aging as a disease as opposed to a normal and even desirable condition? condition? Without Without disease, how would people die? Would older people work forever occupying the jobs potentially available to younger people or be supported in perpetuity by younger generations? Would births cease because the people who live forever take up all the room and eat all the food? According According to Cornellian Kurt Vonnegut, in the future… http://www.tencapitol.com/portfolio_case_phrma1.html 12 Fourteen Painless Ways to Die “All serious diseases had been conquered. So death was voluntary, and the government, to encourage volunteers for death, set up a purple-roofed Ethical roofed Ethical Suicide Suicide Parlor at every major intersection, right next door to an orange-roofed orangeHoward Johnson’s. There were pretty hostesses in the parlor, and Barca-Loungers, Barcaand Muzak, and a choice of fourteen painless ways to die.” If You Advertise it, They will Need it Should Should we have a rational policy to treat some diseases but not others? How How is the sciencesciencepharmaceuticalpharmaceutical-advertising complex redefining what we complex redefining what we consider consider a treatable disease? Is Is healthcare becoming so expensive because more people are living off diseases than are dying from them? No No technology is either a panacea or an Armageddon, but has both advantages and disadvantages. Domestic Applications of Genetic Engineering: Laundry Detergents Enzymes, made by genetic engineering make it possible to get clothes cleaner in cold water, thus saving energy and reducing the output of CO2. http://www.novozymes.com/en/MainStructure/Sustainability/Themes/Environmental+assesment/Intro.htm http://www.novozymes.com/en/MainStructure/Sustainability/Themes/Environmental+assesment/Intro.htm Environmental Applications of Genetic Engineering Bacteria Bacteria with added genes can be used to clean up environmental toxins (e.g. can be used to convert oil or an used to convert oil or an environmental environmental estrogen to carbon dioxide and water). Various Various designer bacteria are matched to efficiently clean up spills of specific chemicals. http://www.novozymes.com/en/MainStructure/ProductsAndSolutions/Bioaugmentation++bioremediation/Surface+cleanup++remediation/BI-CHEM+ABR+Gasoline+Blend/ABR+Gasoline+Blend.htm Enzymes Used to Make Biofuels Cellulosic Ethanol Microbes Microbes can ferment sugars (you know the pathway) into ethanol. A billion tons of sugar a year exists in the form of cellulose, a billion cellulose form of polymerized glucose. It comes from the straw polymerized (inedible stems) of wheat, oats, barley and other grasses. Enzymes Enzymes called cellulases, which are somewhat similar to cellulases amylases amylases, are necessary to convert the unavailable glucose in the form of cellulose into available glucose. Biotech Biotech companies are working on producing efficient enzymes that will convert cellulose to glucose. iogen.ca/cellulose_ethanol www.purevisiontechnology.com/ 13 Cornell Biofuels Research Lab Officially Opens (June, 2009) Plants and Plant Cells as Green Pharmaceutical Factories Carbon dioxide, water and soil nutrients are used by plants to produce vaccines using the energy of sunlight. Vaccines are then isolated from the greenhouse-grown plants. Joyce Van Eck: GeneticallyGeneticallyengineered cultured tomato cells produce a vaccine for poultry against the Deadly Newcastle Disease Virus GeneticallyGenetically-engineered plants and cultured tomato cells produce a vaccine (a vaccine harmless variant of a virus). The genetically-engineered geneticallycultured tomato cells contain the gene that produces a harmless harmless viral protein which acts as an antigen against antigen which the poultry will make antibodies antibodies. Birds inoculated with the vaccine will be resistant to the disease. Transformed Animals also Become Pharmaceutical Factories Transgenic Transgenic Pigs Ensure Uniform and Meaty Pigs Sheep Sheep produce human Erythopoietin. Erythopoietin. 14 From Biotechnology to Chemistry: PostPost-biotic Evolution? The The human hormone secretin secretin is now being synthesized by machines without the need for transcription or translation transcription or translation in in living organisms. It It is being used to diagnose pancreatic exocrine function but could be used for people whose small intestines do not produce secretin. www.chirhoclin.com How do we find the DNA sequence in a human that codes for a particular trait? If If the amino acid sequence of a protein is already known, it is easy to predict the DNA it is easy to predict the DNA sequence. Michael Michael Smith found that he could synthesize a short strand of DNA made of 9-17 9nucleotides that code for the end of the gene he wanted and “fish out” the whole gene using DNADNA-DNA hybridization. DNA Hybridization Test Many Expressed Genes (cDNA) at a Time Using a a Gene Chip or Microarray Test a Whole Human Genome (30,000 Genes) Using A Single Chip the Size of a Dime FISH FISH (Fluorescence In Situ In Hybridization) A fluorescent dye can be attached to the gene probe so th th that the gene can be localized in in situ; that is, in its natural location on the chromosomes using a light microscope. 15 SKY: Spectral KarYotyping Identifying a Single Gene on Chromosome 11 Identify Chromosomal Abnormalities (or Evolutionary Starting Points?) Identify Genes in Embryo Using PrePreImplantation Genetic Diagnosis (PGD) http://www.coloradospringsivf.com/ Polymerase Chain Reaction (PCR) Once Once a gene sequence is identified, we can use polymerase polymerase chain reaction (PCR) to produce many copies (i.e. (i.e. clone) of the gene automatically without the need for living organisms. Kary Kary Mullis invented PCR to PCR clone genes efficiently in vitro. in On On the day he invented PCR… “…Jennifer, “…Jennifer, that crazy, wonderful woman chemist, had dramatically left our house…. I was beginning to learn tragedy…. It would add strength to my character and depth someday to my writing. Just right then, I would have preferred a warm friend to cook with. Hold the tragedy lessons. December is a rotten month to be studying your love life from a distance. I celebrated my victory with Fred Faloona….Fred had helped celebrated me that afternoon set up this first successful PCR this reaction.…I informed him that we had just changed the rules in molecular biology. ‘Okay, Doc, if you say so.’ He knew I was more concerned with my life than with those cute little purplepurpletopped tubes. In In Berkeley it drizzles in the winter. Avocados ripen at odd times and the tree in Fred's front yard was wet and sagging from a load of fruit. I was sagging as I walked out to my little silver Honda Civic…. Neither Fred, empty Becks bottles, nor the the sweet smell of the dawn of the age of PCR could replace Jenny. I was lonesome.” 16 Polymerase Chain Reaction (PCR) Denaturation Denaturation (94 °C) Heat Heat breaks the bonds between the complementary base pairs of the double helix and the two antiparallel strands separate. Roche Roche Molecular Diagnostics Global Holds the Patent for PCR Annealing Annealing (65 °C) Two Two different primers are added at a lower primers temperature so that they can anneal to the DNA. One primer is complementary to the beginning of gene primer is complementary to the beginning of a gene sequence sequence on one strand and the other is complementary to the end of the sequence on the other strand. The four nucleotides containing A, T, C, G are added. The bases of the nucleotides pair with their complementary bases strands on the DNA. Extension (72° Extension (72°C) The DNA The DNA polymerase links together the primers and the annealed nucleotides to make two new strands of DNA, one complementary to each parent strand. http://molecular.roche.com/roche_pcr/index.html DNA polymerase from E. coli E. Initially, Initially, the polymerase chain reactions were done using DNA DNA polymerase from E. coli, E. the bacterium that lives in our large intestine (colon) our large intestine (colon) at at 37 °C. Every Every time the temperature of the DNA was raised to denature the DNA, the enzyme was destroyed and they had to add fresh enzyme during each extension step. Thermus aquaticus: Thermophile Thermus aquaticus lives in hot (70 °C) springs in Yellowstone National Park. In order to live in hot springs, its polymerase must be polymerase must be stable stable at high temperatures, a quality needed for PCR. Cetus Corporation collected the bacterium from this source and isolated Taq Taq polymerase from the bacterium. Everyday Applications of PCR Diagnosis Diagnosis of Genetic Disorders Preimplantation Preimplantation genetic diagnosis Prenatal Prenatal testing Tests Tests for infectious diseases like HIV, hepatitis, Human Papilloma Virus (HPV), etc. in people, animals, the blood supply and the food supply Forensic Forensic Science Innocence Innocence Project Paternity Paternity Testing 17 Diagnoses of Genetic Disorders After in vitro fertilization, in DNA from a single embryonic cell is copied by PCR to diagnose genetic disorders, including including cystic fibrosis, thalassemia, Tay-Sachs Taydisease, Duchenne muscular dystrophy, spinal muscular atrophy, sickle cell anemia, early onset Alzheimer’s disease, hemophilia, and Huntington’s chorea. Preimplantation Genetic Diagnosis On day 3 of embryo development, one or two cells are removed from each embryo and on day 4, the DNA from a cell of each embryo is subjected to PCR using the primers that recognize the gene that encodes the disease of interest. On On day 5, the couple and staff of the clinic use results of the genetic testing to determine which embryo should be transferred into the uterus. PCR is Used for Ensuring a Safe Blood Supply PCR is Used for Testing for HIV, Hepatitis and other Infectious Diseases PCR is Used for Testing the Safety of the Food Supply (e.g. Bird Flu) PCR is Used in Forensic Science to Help Capture Criminals Use Use blood, tissue, semen from a crime scene Extract Extract DNA Perform PCR Compare Compare genetic profile of suspect and “controls” with the genetic profile found on evidence at the crime scene. Present Present evidence to a (scientifically literate) jury 18 Karl Popper: Scientific Theories must be Falsifiable • According to Karl Popper, a universal theory is scientific only if it is falsifiable. • A scientific test can only show that th a hypothesis is false, it cannot prove it to be true. • If the hypothesis is that someone is guilty, DNA evidence can exclude that person as the guilty party but it cannot verify that the person is guilty. Innocence Project: Barry Scheck and Peter Neufeld Use PCR to Set Innocent People Free PCR is Used for Paternity Testing: A Pop Quiz? Based on this One Locus, this Man Cannot Be Excluded as the Possible Father The DNA markers from the mother and the putative father account for the DNA markers of for the DNA markers of the child at this locus. The father’s DNA at this locus accounts for all of the child’s DNA at this locus which could not have come from the mother. 19 Alleles from Many Loci are Tested Alleles Alleles from loci 1,5, 7 and 10 do not exclude the do putative father. Alleles Alleles from loci 2, 3, 4, 6, 8 and 9 do exclude the exclude the putative putative father. If If a putative father cannot be excluded, the likeliness likeliness of paternity is calculated by taking into consideration the number of loci tested and the frequencies of the alleles at the various loci. “Home” Paternity Tests http://www.dnacenter.com/ idna-systems.com Paternity Testing In lab you are going to perform PCR on your own DNA and get your genetic profile for the D1S80 locus on Chromosome 1 bio-rad.cnpg.com Rejection of PCR Manuscript “I “I knew that PCR would spread across the world like wildfire. This time there was no doubt in my mind: Nature would publish Nature it. They rejected it. So did Science Science, the second most prestigious journal in the world. Science Science offered that perhaps my paper could be published in some secondary journal, as they felt it would not be suitable to the needs of their readers, ‘F*** them,’ I said.” Kary Mullis and PCR “It was some time before my disgust with the journals mellowed. I accepted an offer by Ray Wu to publish it in Methods of Enzymology, a volume he Methods was preparing. He understood the power of PCR.” 20 “His book challenges us to question the authority of scientific dogma even as it reveals the workings workings of an uncannily original scientific mind.” http://www.karymullis.com/ Marketing DNA Individuality 1. Swab the inside of your cheek, mail sample to lab. 2. Your sample is processed into a DNA profile. 3. Your fragrance is formulated and mailed to you. www.mydnafragrance.com Personalized DNA Art Personalized DNA Art http://www.dna11.com/ 21 Personalized DNA Art Your Choice There There is no class on Friday, March 19 (Dragon Day). On On Monday, March 29 at 9:05 can give at 9:05, I can give a lecture lecture on “What I did over Spring break” or we all can sleep late. Let’s Let’s take a vote. All All in favor say, “Aye”. Opposed? Opposed? Have a Fun, Healthy and Safe Spring Break! http://www.happyhourcomedy.com/ http://happyhourcomedy.com/blog/about/ http://happyhourcomedy.com/blog/ 22 ...
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