54. DNA techLec10 - DNATechnology DNA Technology lab...

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1 DNA Technology DNA Technology lab sequence Week 1: Cloning of gene or genetic marker Week 2: Electrophoresis; Restriction mapping Week 3: DNA sequencing; PCR of human microsatellite DNA Week 4: Analysis of your own genotype; forensic application of human microsatellite DNA Studying how gene functions or to learn its nucleotide sequence often requires: ! Isolation of single gene away from other genes in the genome ! Methods to produce large quantity of the gene so that its function can be observed easily Restriction Enzyme Structure - Most restriction enzymes are homodimers GGATTC CTTAGG Bam H1 Restriction Enzyme - Restriction enzymes are specific enzymes in bacteria that cut foreign DNA for defense - After being “restricted”, the foreign DNA can be digested by exonucleases, enzymes that digest DNA from ends one base at a time - In contrast, restriction enzymes are also called endonucleases endonucleases exonucleases - Restriction enzymes recognize specific sequences of DNA and cut both strands in specific ways - The sequences recognized are 4 to 8 base pairs - The sequences are palindromes - Restriction reaction creates reproducible DNA segments called restriction fragments Restriction Enzyme Mechanism GAATTC CTTAAG
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2 Restriction Enzyme Cutting - Some RE cut create blunt ends
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This note was uploaded on 03/13/2010 for the course BIOG 1102 taught by Professor Robertturgeo during the Spring '10 term at Cornell University (Engineering School).

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54. DNA techLec10 - DNATechnology DNA Technology lab...

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