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IV Manipulating DNA The Tools of Molecular Biology The Tools of Molecular Biology How do scientists make changes to DNA? The Tools of Molecular Biology Scientists use different techniques to: • extract DNA from cells • cut DNA into smaller pieces • identify the sequence of bases in a DNA molecule • make unlimited copies of DNA The Tools of Molecular Biology In genetic engineering, biologists make changes in the DNA code of a living organism. 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. Which type of molecule is an enzyme? 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 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. 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 DNA plus restriction enzyme First, restriction enzymes cut DNA into fragments. The DNA fragments are Mixture of DNA poured into wellsfragments on a gel. Gel Electrophoresis Gel The Tools of Molecular Biology Power source An electric voltage is applied to the gel. The smaller the DNA fragment, the faster and farther it will move across the gel. The Tools of Molecular Biology Power source Longer fragments Shorter fragments Gel Electrophoresis Using the DNA Sequence Making Copies Polymerase chain reaction (PCR) is a technique that allows biologists to make copies of genes. Small amounts of DNA can be multiplied making it easier to analyze. Made possible by an enzyme found in a bacterium living in hot springs in Yellow Stone National Park. 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. Quiz Restriction enzymes are used to • • • • extract DNA. cut DNA. separate DNA. replicate DNA. During gel electrophoresis, the smaller the DNA fragment is, the • • • • more slowly it moves. heavier it is. more quickly it moves. darker it stains. The DNA polymerase enzyme found in bacteria living in the hot springs of Yellowstone National Park illustrates • genetic engineering. • the importance of biodiversity to biotechnology. • the polymerase chain reaction. • selective breeding. A particular restriction enzyme is used to • • • • cut up DNA in random locations. cut DNA at a specific nucleotide sequence. extract DNA from cells. separate negatively charged DNA molecules. During gel electrophoresis, DNA fragments become separated because • • • • multiple copies of DNA are made. recombinant DNA is formed. DNA molecules are negatively charged. smaller DNA molecules move faster than larger fragments. END OF SECTION