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Chapter 13 Genetic Engineering Section 13-2 Manipulating DNA FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Manipulating DNA Key Concept: Scientists Use Their Knowledge Of The Structure of DNA And Its Chemical Properties To Study and Make Changes To DNA Molecules FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Manipulating DNA Key Concept (cont.) Different Techniques are used to Extract DNA from Cells, to Cut DNA into Smaller Pieces, to Identify the Sequence of Bases in the DNA Molecule, and to Make Unlimited Copies of DNA FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT The Tools of Molecular Biology Genetic Engineering The Process of Making Changes In The DNA Code of a Living Organism FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT The Tools of Molecular Biology DNA Extraction Use Chemicals To Lyse Cells Lysis: a process of disintegration or dissolution (as of cells) FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT The Tools of Molecular Biology Cutting DNA DNA Too Large To Work With, So It Is Cut Up Using: Restriction Enzymes: • They Are Very Precise • 100’s Are Known • Each One Cuts DNA At Specific Sequence of Nucleotides FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Restriction Enzymes FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT The Tools of Molecular Biology Separating DNA Gel Electrophoresis • Mixture of Fragments Put In One End of Gel • Electric Voltage Applied • Fragments Travel Toward Positive End of Gel FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT The Tools of Molecular Biology Gel Electrophoresis (cont.) –Smaller The Fragment, The Faster It Moves –Used To: • Compare Genomes • Locate Individual Genes • Identify Base Pair Sequence FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Gel Electrophoresis FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Gel Electrophoresis FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Gel Electrophoresis FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Gel Electrophoresis FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence Once The DNA Is A Manageable Size, You Can: 1. Read The Nucleotide Sequences 2. Modify The Genome FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence Reading The Sequences – Now Automated – Small, Single Strands of DNA – Add Enzyme That Makes Complementary Strand – Add Nucleotides, Some That Are Labeled With A Specific Color of Fluorescent Dye FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence Reading The Sequences (cont.) –Addition of a Labeled Base To Strand Terminates Replication –Produces Multiple, Labeled Strands of Different Lengths –Each Terminal Base Is Color Coded FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence Reading The Sequences (cont.) –Separate by Electrophoresis –Read The Base Sequences In Order, By Color Codes FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Automated Sequencing FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Changing The DNA Sequence Cutting & Pasting You Can Take A Gene From One Organism & Attach It To The Gene of Another Organism FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Changing The DNA Sequence Cutting & Pasting (cont.) Mixing Genes From Different Organisms Results In Recombinant DNA FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence Making Copies PCR = Polymerase Chain Reaction Quickly Makes Multiple Copies Of Small DNA Targets FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT PCR Polymerase Chain Reaction Add Short Strand of Complementary DNA To Each End Of The DNA You Want To Copy Primers Provide Location For DNA Polymerase To Attach FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT PCR Polymerase Chain Reaction • Heat To Separate The Strands Of DNA • Cool To Allow Primers To Attach The Primers • Increase Temperature To Activate TAQ Polymerase (High Temperature DNA Polymerase) • Repeat 20 – 30 Times – Copies Also Act As Templates FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT PCR Polymerase Chain Reaction Within A Few Hours You Will Have Millions of Copies FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT PCR Polymerase Chain Reaction FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT PCR Polymerase Chain Reaction • Kary Mullis – Invented PCR – Source of High Temperature DNA Polymerase (taq Polymerase) • Bacteria In hot springs of Yellowstone Park FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT PCR Polymerase Chain Reaction Yellowstone Hotspring Source of High Temperature PCR DNA Polymerase (taq Polymerase) FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT