Team:Osaka/week3

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Revision as of 06:22, 18 August 2012 by Toty (Talk | contribs)

August 12 (Sun)

  1. Miniprep of yesterday's culture
  2. Restriction digests of minipreppped parts
  3. Gel electrophoresis of digests
  4. Ligation
    • 153(C):1-2M(upstream)+1-10H(downstream)+1-1A(vector)
    • 154(C):1-22A(upstream)+1-2M(downstream)+1-1A(vector)
    • 155(C):1-2D(upstream)+1-23L(downstream)+1-1A(vector)
  5. SDS-PAGE
    • Gel electrophoresis

August 16 (Thu)

  1. Transformaton of ligation products into DH5α E.coli:153~155
  2. Transformation of plasmid DNA into Rosetta E.coli:empty vector(A)

August 17 (Fri)

  1. Transfer to liquid culture

August 18 (Sat)

  1. Preparation of LB agar plates (Tet,Kan)
  2. Miniprep of yesterday's culture
  3. PCR
    • mutation was found in PprA → retrial
  4. Purification of PCR products
  5. Restriction digests of minipreppped parts
  6. Gel electrophoresis of digests
    • 155 x
  7. Ligation
    • 156(T):153(upstream)+1-23L(downstream)+1-7A(vector)
  8. Transformation of ligation products into DH5α E.coli:155,156

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