week 9/16 to 9/20

  • Gram-stained Sonora plates from 9/13
    • short rods 
    • Gram Positive
  • plasmid extraction of Cas9 
  • Gel excision prep
    • cut wanted band from gel 
    • For every actual weight of agarose gel, add 100 µL of binding buffer. 
    • Place in incubator until gel is dissolved (should be yellow if buffer is working). 
    • Transfer 400 µL of gel solution to a microcentrifuge tube for 1 min, then discard the flowthrough 
    • 100 µL of binding buffer for 1 min, then discard flowthrough. 
    • 100 µL of wash buffer, then centrifuge for 1 min to discard flowthrough
    • Then place the column in a new centrifuge tube to add 20 µL of elution buffer.
  • learned that E. coli is what is used to keep bacteria
  • restriction sites of PGRNA is about 6 bp
    • cut a site to make linear 
  • C1. V1 = C2. V2 
    • C 1 starting  concentration 
    • V1, starting volume 
    • C2, ending concentration 
    • V2 ending volume 

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week 10/28- 11/1

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week 10/21-25

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