FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT 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 In The DNA Code of a Living Organism 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 PreciseThey Are Very Precise 100’s Are Known100’s Are Known Each One Cuts DNA At Specific Sequence of NucleotidesEach 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 GelMixture of Fragments Put In One End of Gel Electric Voltage AppliedElectric Voltage Applied Fragments Travel Toward Positive End of GelFragments 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 GenomesCompare Genomes Locate Individual GenesLocate Individual Genes Identify Base Pair SequenceIdentify 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

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 Recombinant DNARecombinant DNARecombinant DNA

FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Using The DNA Sequence Making Copies PCR = Polymerase Chain Reaction 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 DNAHeat To Separate The Strands Of DNA Cool To Allow Primers To Attach The PrimersCool To Allow Primers To Attach The Primers Increase Temperature To Activate TAQ Polymerase (High Temperature DNA Polymerase)Increase Temperature To Activate TAQ Polymerase (High Temperature DNA Polymerase) Repeat 20 – 30 TimesRepeat 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 MullisKary Mullis –Invented PCR –Source of High Temperature DNA Polymerase (taq Polymerase) Bacteria In hot springs of Yellowstone ParkBacteria 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)