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Amgen Lab 2a & 4a

Amgen Lab 2a & 4a. Lab 2a: Restriction Enzymes. Restriction enzymes are naturally found in Bacteria & Archaea . They are thought to provide protection against invading viruses by hydrolyzing “foreign” DNA. RE cut DNA at specific recognition nucleotide sequences called restriction sites

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Amgen Lab 2a & 4a

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  1. Amgen Lab 2a & 4a

  2. Lab 2a: Restriction Enzymes • Restriction enzymes are naturally found in Bacteria & Archaea . • They are thought to provide protection against invading viruses by hydrolyzing “foreign” DNA. • RE cut DNA at specific recognition nucleotide sequences called restriction sites • Called “molecular scissors”

  3. Produces “sticky ends”

  4. BamH I sticky end Hind III sticky end Hind III sticky end BamH I sticky end Engineering the Plasmid: ligation of rfp gene into p-ARA Bruce Wallace

  5. pARA-R construct Bruce Wallace Recombinant plasmid of interest pARA-R 4720 bp BamH I rfp 702bp Hind III

  6. Restriction digest of pARA-R Bruce Wallace Recombinant plasmid of interest pARA-R 4720 bp rfp 702bp BamH I Hind III

  7. Restriction analysis of pARA-R Bruce Wallace Restriction fragments after digest with Hind III and BamH I BamH I Hind III 4018 bp BamH I Hind III 702bp

  8. 10000 8000 5000 4000 3000 2000 1500 1000 500 Restriction analysis of pARA-R Prediction for restriction gel A- A+ M A- A+ M

  9. A- Lane: Nicked, Multimer, & “Regular” plasmid

  10. Ladder A+ A-

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