Team:Osaka/week8

September 16 (Sun)

1. Miniprep of yesterday's culture
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. Transfer to liquid culture
8. Tolerance assay(140,150)

September 17 (Mon)

1. Miniprep of yesterday's culture:161,162
2. Restriction digests of minipreppped parts
3. Transfer to liquid culture:163~166,
4. Electrophoresis of digests
5. Tolerance assay(WT)

September 18 (Tue)

1. Miniprep of yesterday's culture:163~166
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
4. Transfer to liquid culture

September 19 (Wed)

1. Gel extraction :161,162
2. Transfer to liquid culture:161,162
3. Miniprep of yesterday's culture
4. Restriction digests of minipreppped parts
5. Electrophoresis of digests
6. Transformation of plasmid DNA into E.coliDH5α 163~166 / HST04 161,162

September 20 (Thu)

1. Miniprep of yesterday's culture:164~166
2. Restriction digests of minipreppped parts
3. Electrophoresis of digests
4. Tolerance assay(149,150,152,WT)
5. Sequencing (160~162,165,166)
6. Restriction digests(1-1D)
7. Ligation
   • 167(C):1-1D(upstream)+160(downstream)+1-3A(vector)
8. Transformation of ligation products into DH5α E.coli
9. Transformation of plasmid DNA into E.coliHST04 165,166
10. Preparation of glycerol stock of cell culture:161~166
11. Restriction digests
12. Ligation
    • 168(T):1-1D(upstream)+153(downstream)+1-7A(vector)
    • 169(T):159upstream)+153(downstream)+1-7A(vector)
    • 170(T):1-9N(upstream)+153(downstream)+1-7A(vector)
    • 171(T):1-23L(upstream)+154(downstream)+1-7A(vector)
    • 172(C):1-2D(upstream)+1-23L(downstream)+1-3A(vector)
13. Transformation of ligation products into DH5α E.coli

September 21 (Fri)

1. Miniprep of yesterday's culture:163,164
2. Restriction digests of minipreped parts
3. Electrophoresis of digests
4. Tolerance assay:143~146,150
5. To liquid culture:WT,147,148,161~166

September 22 (Sat)

1. Miniprep of yesterday's culture:163,164,167
2. Restriction digests of minipreped parts
3. Electrophoresis of digests
4. Tolerance assay:146~148,WT
5. Transformation of 01,02,03,04,167 into HST04 E.coli

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