Genetic Engineering and Biotechnology Notes IB Assessment Statement 4.4.1 Outline the use of polymerase chain reaction (PCR) to copy and amplify minute quantities of DNA. Copyright Pearson Prentice Hall The Tools of Molecular Biology Scientists use different techniques to: • make unlimited copies of DNA • extract DNA from cells • cut DNA into smaller pieces • identify the sequence of bases in a DNA molecule Using the DNA Sequence Making Unlimited Copies • Polymerase chain reaction (PCR) is a technique that allows biologists to make copies of genes. • It is useful when small amounts of DNA are found (Example: at a crime scene) Using the DNA Sequence DNA is heated to separate its two strands, then cooled to allow the primers to bind to singlestranded DNA. DNA polymerase enzyme starts making copies of the region between the primers. The DNA Polymerase Enzyme is removed from a bacteria Thermus aquaticus Using the DNA Sequence Polymerase Chain Reaction (PCR) DNA heated to separate strands DNA polymerase adds complementary strand DNA fragment to be copied PCR cycles 1 DNA copies 1 2 2 3 4 4 8 5 etc. 16 etc. PCR ANIMATION http://highered.mcgrawhill.com/olc/dl/120078/micro15.swf http://www.sumanasinc.com/webcontent/animations/c ontent/pcr.html Copyright Pearson Prentice Hall PCR Virtual Lab – Upload answers on Moodle Click here to go to Virtual Lab Website: http://learn.genetics.utah.edu/content/labs/pcr/ Answer the below questions: 1.What is the overall purpose of PCR? 2.What does the why is a primer, DNA polymerase, and nucleotides necessary for the reaction? 3.Why must you lower and raise the temperature of the reaction? Copyright Pearson Prentice Hall IB Assessment 4.4.2 State that, in gel electrophoresis, fragments of DNA move in an electric field and are separated according to their size. Copyright Pearson Prentice Hall The Tools of Molecular Biology DNA Extraction DNA can be extracted from most cells by a simple chemical procedure. The cells are opened and the DNA is separated from the other cell parts. The Tools of Molecular Biology Cutting DNA Most DNA molecules are too large to be analyzed, so biologists cut them into smaller fragments using restriction enzymes. The Tools of Molecular Biology Each restriction enzyme cuts DNA at a specific sequence of nucleotides. Recognition sequences DNA sequence Restriction enzyme EcoR I cuts the DNA into fragments Sticky end The Tools of Molecular Biology A restriction enzyme will cut a DNA sequence only if it matches the sequence precisely. Recognition sequences DNA sequence Restriction enzyme EcoR I cuts the DNA into fragments Sticky end The Tools of Molecular Biology Separating DNA In gel electrophoresis, DNA fragments are placed at one end of a porous gel, and an electric voltage is applied to the gel. When the power is turned on, the negativelycharged DNA molecules move toward the positive end of the gel. The Tools of Molecular Biology Gel electrophoresis can be used to compare the genomes of different organisms or different individuals. It can also be used to locate and identify one particular gene in an individual's genome. The Tools of Molecular Biology Power source DNA plus restriction enzyme Longer fragments Mixture of DNA fragments Gel Gel Electrophoresis Shorter fragments The Tools of Molecular Biology First, restriction enzymes cut DNA into fragments. DNA plus restriction enzyme The DNA fragments are poured into wells on a gel. Mixture of DNA fragments Gel Electrophoresis Gel The Tools of Molecular Biology An electric voltage is applied to the gel. This moves the DNA fragments across the gel. The smaller the DNA fragment, the faster and farther it will move across the gel. Power source Gel Electrophoresis The Tools of Molecular Biology Based on size, the DNA fragments make a pattern of bands on the gel. These bands can then be compared with other samples of DNA. Longer fragments Shorter fragments Gel Electrophoresis IB ASSESSMENT STATEMENT 4.4.3 State that gel electrophoresis of DNA is used in DNA profiling. Copyright Pearson Prentice Hall IB Assessment Statement 4.4.4 Describe the application of DNA profiling to determine paternity and also in forensic investigations. 4.4.5 Analyse DNA profiles to draw conclusions about paternity or forensic investigations. Copyright Pearson Prentice Hall This DNA fingerprints are used to do DNA Profiling. There are two many uses for DNA profiling: 1. Forensic use – determine the guilt of a criminal 2. Paternity Test – determine who the father of an offspring IB Assessment Statement 4.4.4 Describe the application of DNA profiling to determine paternity and also in forensic investigations. 4.4.5 Analyse DNA profiles to draw conclusions about paternity or forensic investigations. Copyright Pearson Prentice Hall Gel Electrophoresis Lab Click here to go to the website: http://learn.genetics.utah.edu/content/labs/gel/ Answer the questions and address the tasksbelow: 1.Summarize the procedure of gel electrophoresis 2.State two properties of the fragmented pieces of DNA which allow them to be separated in gel electrophoresis. Copyright Pearson Prentice Hall This DNA fingerprints are used to do DNA Profiling. There are two many uses for DNA profiling: 1. Forensic use – determine the guilt of a criminal 2. Paternity Test – determine who the father of an offspring Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Click here to go the website: http://learn.genetics.utah.edu/archive/mystery/records.html The Case of the Pot Hunters Click here to go the website: http://learn.genetics.utah.edu/archive/mystery/records.html Then read the prepared evidence , analyze the gel electrophoresis results, and read the courts transcript, and you decide whether or not the suspect is guilty. Write your decision on an MS Word document. Be sure to clearly state whether you think the suspect is gulity/ not guilty/ or if there is not enough information to say either way. Support your statement with scientific evidence and logical reasoning. Copyright Pearson Prentice Hall IB Assessment Statement Outline three outcomes of the sequencing of the complete human genome. Copyright Pearson Prentice Hall Outline three outcomes of the sequencing of the complete human genome. Copyright Pearson Prentice Hall Human Genome Project • A project set out to sequence the complete human genome. • It hoped to determine all the bases A, T, C, G in the human DNA. • In 2003, it succeeded. • Now scientists are trying to figure out which base sequence represents genes. Copyright Pearson Prentice Hall Human Genome Project: It can be thought of as a map showing the locus of any gene on the 23 pairs of chromosomes. Copyright Pearson Prentice Hall Copyright Pearson Prentice Hall Benefits of the Human Genome Project: • Find out if people are carrying a disease causing allele • Find beneficial molecules which are produced naturally by healthy people • Find out which gene controls the synthesis of a desirable molecule • Copy the above gene and use t as instructions to synthesize the molecule in a laboratory • Distribute above molecule as a new medical treatment Copyright Pearson Prentice Hall Human Genome • It is estimated that humans consist of 25,000-30,000 genes. • More the 1500 genes have been identified that cause disease. Copyright Pearson Prentice Hall http://www.youtube.com/watch?v=zi8FfMBYCkk Copyright Pearson Prentice Hall Outline three outcomes of the sequencing of the complete human genome. Copyright Pearson Prentice Hall Human Genome Project Click on the below website and watch the INTRODUCTION animation. In your own words explain what the Human Genome Project is: http://www.genome.gov/25019885 Copyright Pearson Prentice Hall http://www.hhmi.org/biointeractive/m edia/DNAi_genetic_eng-sm.mov http://www.ted.com/talks/lang/en/pau l_root_wolpe_it_s_time_to_question _bio_engineering.html