LECTURE 17 Biotech S10

LECTURE 17 Biotech S10 - Announcements CAPE: Due online!...

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1 Announcements Final Exam Preparation - THIS IS IT! PRACTICE FINAL EXAM posted on website EXTRA OFFICE HOURS this /some next week (see website) REVIEW SESSION Wed. June 9th, 6:00-9:00 PM, Ledden 216 FINAL EXAM Fri. June 12th, 8:00-11:00 AM, Solis 107 (the mothership) Comprehensive Exam Will cover new and old material (Chpt. 2-20; not Chpt. 11) GOOD LUCK! CAPE: Due online! Please share your opinion. Only 20.73% response rate so far…(Other BILD1 class is at 33.82%!) Lecture 17 DNA Technology Understanding and Manipulating Genomes DNA Technology is application of learned properties of DNA The Molecular Revolution launched advances in: - Biological research human disease, mechanisms of development, evolutionary underpinnings - Biotechnology manipulation of organisms or components to make products Applications of Biotechnology Disease diagnostics Disease treatments Forensic science Agriculture Environmental applications Clone your favorite pet … …?
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2 DNA Technology I. Amplifcation oF DNA A. DNA Cloning B. PCR II. Detection oF DNA A. Gel electrophoresis B. Sequencing C. Hybridization III. Signifcance Understand the basic concepts and components oF these techniques I. Amplifcation oF DNA DNA amplifcation to generate multiple copies oF a specifc gene or DNA segment Since specifc gene sequences usually represented once , thereFore only small Fraction oF chromosomal (genomic) DNA A. Cloning - amplifcation by replication inside cells B. PCR - amplifcation by purifed enzymes outside cells AMPLI±ICATION A. DNA Cloning Production oF multiple copies oF a DNA segment Use oF bacterial plasmids (vectors) to carry DNA Bacteria are used as hosts to replicate copies oF DNA DNA Cloning using bacterial plasmids Figure 20.2 Piece oF human DNA Bacterial plasmid Bacterial reproduction results in replicated copies oF human DNA on plasmid DNA Protein application
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3 Restriction Enzymes & Ligase are used to make recombinant DNA Two important frst steps in Cloning require two enzymes 1. Restriction Enzymes – Bacterial enzymes that cut at specifc restriction site sequences – Cut DNA by breaking phosphodiester bonds – Yield restriction Fragments that can be used to clone by complementary ends 2. DNA Ligase - Enzyme reForms phosphodiester bond between adjacent nucleotides ±igure 20.3 Restriction site DNA 5’ 3’ 5’ 3’ G A A T T C C T T A A G “Sticky ends” G C T T A A A A T T C G G G G G A A T T C A A T T C C T T A A G C T T A A G Recombinant DNA Cloning Using a restriction enzyme and DNA ligase to make recombinant DNA Restriction enzyme Ligase Green gene (±ragment 1) + Grey gene (±ragment 2) = Recombinant green-grey gene e.g. EcoRI enzyme cuts at “GAATTC” palindrome Using a Bacterial Plasmid to Clone a Eukaryotic Gene • A bacterial plasmid can serve as a cloning vector
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This note was uploaded on 10/16/2010 for the course BILD 1 taught by Professor Boulanger during the Spring '08 term at UCSD.

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LECTURE 17 Biotech S10 - Announcements CAPE: Due online!...

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