Team:Osaka/week3

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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