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Which sizes of DNA fragments do you expect to get from the digestion?

Journal 12.09.10:. Which sizes of DNA fragments do you expect to get from the digestion? 2. How can one use the electrophoresis procedure to estimate the size of the DNA fragments?. pARA-R construct. Recombinant plasmid of interest. pARA-R 4720 bp. pARA-R. 4720 bp. Bam H I. rfp 702bp.

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Which sizes of DNA fragments do you expect to get from the digestion?

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  1. Journal 12.09.10: • Which sizes of DNA fragments do you expect to get from the digestion? • 2. How can one use the electrophoresis procedure to estimate the size of the DNA fragments?

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

  3. Gel Electrophoresis - Detection We run the gel until the yellow dye runs out. Wells

  4. After running the gel (till the yellow dye runs out), we stain it in Ethidium bromide, which will fluoresce when bound to DNA:

  5. Supercoiling Multimers “nicked-circle” circle

  6. Structural forms of plasmid Slower than nicked. “multimer” “nicked-circle” Linear (Digested) Supercoiled

  7. A- A+ 10 Kb Ladder 10 Kb Ladder 10 Kb Ladder Multimer Nicked Super Coiled 5 Kb Linear Fragment Linear Fragment

  8. M A- A+ Yellow highlighter shows group number.. Expected results

  9. Class Results 2013: Lab 4a

  10. Groups 1,2

  11. Groups 3,4

  12. 4000base pairs 2000base pairs 1500 base pairs 1000 base pairs 500 base pairs

  13. Lab 2a (restriction) and 4a (electrophoresis) almost always come together – why is that? • 2) Point out at least one thing that you learned so far that is important to know when working in a biotech lab.

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