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Biotechnology

This chapter explores the origins and processes involved in recombinant DNA technology, including restriction enzymes, gel electrophoresis, transformation, and molecular cloning. It also discusses the various vectors used in cloning and DNA libraries, as well as techniques such as hybridization, DNA sequencing, and genetic engineering. Includes an assignment on analyzing the pros and cons of genetic engineering.

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Biotechnology

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  1. Biotechnology Chapter 17

  2. How did it all start? • Recombinant DNA

  3. How is Recombinant DNA made? • Restriction Enzymes • Restriction sites • Type 1 • Type 2 • Dna Ligase

  4. Gel Electrophoresis • Separates fragmented DNA • Takes advantage of DNA’s negative charge • Separates by size

  5. Transformation • So what is it? • In E.coli • Transgenenic Organisms

  6. Molecular Cloning • Host-Vector Systems • Commonly used host? • Commonly used vectors? • How to clone a gene!

  7. Plasmid Vector • Must have • 1. Origin of replication • 2. Selectable marker

  8. Phage Vectors • Can hold more inserted DNA up to 40 kb • Lambda phages kill cells, and are linear

  9. DNA Libraries • DNA Library vs Genomic Library • Reverse Transcriptase • Making cDNA • Hybridization

  10. Steps to Hybridization • 1. Colonies of plasmid w/ bacteria each containing a singe DNA from the library are grown on agar. • 2. A replica of the plate is made using filter paper • 3. Filter paper is washed in a solution that breaks cells open and denatures DNA- the filter is incubated with radioactively labeled probe that will from hybrids with complimentary DNA gene of interest. • 4. Colonies of interest will retain the radioactive probe. • 5. A comparison between the original plate and the filter paper is made. • FIGURE 17.6

  11. DNA ANALYSIS • Restriction Maps • Southern Blots • RFLP Analysis • DNA fingerprinting • PCR

  12. DNA Sequencing • Enzymatic Sequencing • PCR • Applications

  13. Genetic Engineering • ASSIGNMENT!!!!!!!!!!! • Read and Outline IN DEPTH Pages 339-346. • Develop two arguments one for and one against the use of genetic engineering. • Write a 2 page paper outlining both the pro’s and con’s of Genetic Engineering • Be prepared to present your findings in class!!!!!!

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