Team:Osaka/week7

September 9 (Sun)

1. Miniprep of yesterday's culture(150,151,152,159,160)
2. Restriction digests of minipreppped parts
3. Electrophoresis
4. Tolerance assay of 144,147(MMC)
5. Transfer to liquid culture(144,145,160)

September 10 (Mon)

1. Miniprep of yesterday's culture(160,1-9N)
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
4. Preparation of LB agar plates(Amp)
5. Ligation
   • 160(K):157(upstream)+158(downstream)+1-5A(vector)
6. Transformation of ligation products
7. Transfer to liquid culture(146,148)
8. Tolerance assay of 144,145(MMC)

September 11 (Tue)

1. Restriction digests(157,158,159,160,1-9N)
2. Gel electrophoresis of digests
3. Ligation
   • 160(K):157(upstream)+158(downstream)+1-5A(vector)
4. Transformation of ligation products
5. Tolerance assay of 146,148(MMC)

September 12 (Wed)

1. Miniprep of yesterday's culture:160
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
4. Tolerance assay
5. Ligation
   • 161(C):1-9N(upstream)+160(downstream)+1-3A(vector)
   • 162(C):159(upstream)+160(downstream)+1-3A(vector)

September 13 (Thu)

1. Colony check
   • 161:Transformed cells didnot grow
2. Sequencing(157,158,159,160)
3. Tolerance assay of 146,148
4. Transformation of ligation products:161
5. Transfer to liquid culture(144~148,162)

September 14 (Fri)

1. Miniprep of yesterday's culture:162
2. Restriction digests of minipreppped parts
3. Tolerance assay (Preliminary test of hydrogen peroxide)
4. Gel electrophoresis of digests
5. Transfer to liquid culture:149,152,161

September 15 (Sat)

1. Miniprep of yesterday's culture:161
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
4. Gel extraction :161,162
5. Ligation
   • 163(C):161(upstream)+151(downstream)+1-3A(vector)
   • 164(C):161(upstream)+152(downstream)+1-3A(vector)
   • 165(C):162(upstream)+151(downstream)+1-3A(vector)
   • 166(C):162(upstream)+152(downstream)+1-3A(vector)
6. Transformation of ligation products into DH5α E.coli
7. Tolerance assay (Hydrogen peroxide)
8. Transfer to liquid culture:

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