Team:Osaka/week3

From 2012.igem.org


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
    • Electrophoresis

August 16 (Thu)

  1. Transformaton of ligation products into DH5α E.coli:153~155
  2. Transformation of plasmid DNA into Rosetta E.coli:151

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:153~155
  3. PCR
    • mutation was found in PprA → retrial
  4. Purification of PCR products
  5. Preparation of glycerol stock of cell culture:151
  6. Restriction digests of minipreppped parts
  7. Gel electrophoresis of digests
    • 155 x
  8. Ligation
    • 156(T):153(upstream)+1-23L(downstream)+1-7A(vector)
  9. Transformation of ligation products into DH5α E.coli:155,156

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