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Manipulating DNA Manipulating DNA • Scientists use their knowledge of the structure of DNA and its chemical properties to study and change DNA molecules • Different techniques are used to study and change DNA molecules Tools of Molecular Biology • Genetic Engineering – making changes in the DNA code of a living organism • DNA extraction • Cells opened to separate DNA from other cell parts • Cutting DNA – DNA too large to study, so biologists “cut” them into smaller fragments using restriction enzymes. Many restriction enzymes are known and each one cuts DNA at a specific sequence of nucleotides Restriction Enzymes Recognition sequences DNA sequence Restriction enzyme EcoRI cuts the DNA into fragments. Sticky end Restriction Enzymes Recognition sequences DNA sequence Restriction enzyme EcoRI cuts the DNA into fragments. Sticky end Separating DNA • Gel electrophoresis – Used to separate DNA fragments. DNA fragments placed in a gel and electricity is applied to the gel. DNA molecules are negatively charged and move towards the positive end of the gel. Smaller DNA fragments move faster and farther – This technique used to compare the genomes of different organisms or even different people Gel Electrophoresis Power source DNA plus restriction enzyme Longer fragments Shorter fragments Mixture of DNA fragments Gel Gel Electrophoresis Gel Electrophoresis Which suspect should have more questioning? DNA from Victim DNA DNA DNA DNA Blood from from from from found at Suspect Suspect Suspect Suspect Crime #2 #3 #4 #1 Scene Using the DNA Sequence • We can use gel electrophoresis to figure out the DNA sequence. By knowing the DNA sequence, we can study and compare specific genes. Cutting and Pasting DNA • DNA sequences can be changed by splicing original DNA with “synthetic” pieces from another organism • Splicing – adding in a new section of DNA into an existing piece of DNA • Recombinant DNA – combining DNA from different sources – Ex. Taking E. coli and inserting the human gene for making insulin into the bacteria’s DNA so that it produces human insulin for diabetics Making Copies of DNA • Polymerase Chain Reaction (PCR) technique – Allows biologists to make many copies of DNA – DNA strands separated with heat, then cooled to allow DNA Polymerase to start making new copies of DNA – A few dozen heat and cool cycles results in many copies of DNA Making Copies of DNA DNA polymerase adds complementary strand DNA heated to separate strands DNA fragment to be copied PCR cycles 1 2 3 4 5 etc. DNA copies 1 2 4 8 16 etc.