Team:Osaka/week4

Contents 1 August 21(Sun) 2 August 22(Mon) 3 August 23(Tue) 4 August 24(Wed) 5 August 25(Thu) 6 August 26(Fri) 7 August 27(Sat)

August 21(Sun)

1. Preparation of LB agar plates (14Tet)
2. Transfer to liquid culture:01,02,03,04
3. Transformaton of Registry parts (See Table 4).

Table4

ID	Part Name	Resistance	Description
1-7A	<bbpart>BBa_J04450</bbpart>	T	construction plasmid containing mRFP coding device

August 22(Mon)

1. Colony check
   • Transformed cells produced colonies!
   • Non-transformed cells (negative controls) did not grow on Tet plates -> confirmed lack of natural antibiotic resistance
2. Transfer to liquid culture:1-7A

August 23(Tue)

1. Miniprep of yesterday's culture
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
   • 1-7A ok

August 24(Wed)

1. Gel electrophoresis of 1-7A
2. Gel extraction of 1-7A
3. Ligation
   • 08(T):07(upstream)+1-23L(downstream)+1-7A(vector)
4. Transformation of 08 and Registry parts (see Table5)

Table5

ID	Part Name	Resistance	Description
4-15M	<bbpart>BBa_K325101</bbpart>	C	EPIC Firefly Luciferase　Photinus Pyralis　(E. coli optimised)

August 25(Thu)

1. Transfer to liquid culture:08,4-15M

August 26(Fri)

1. Miniprep of yesterday's culture
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
   • 08 ×
   • 4-15M ok
4. Ligation
   • 09(K):1-9N(upstream)+4-15M(downstream)+1-5A(vector)
   • 10(K):1-1D(upstream)+4-15M(downstream)+1-5A(vector)
5. Transfer to liquid culture:08

August 27(Sat)

1. Miniprep of yesterday's culture
2. Restriction digests of minipreppped parts
3. Gel electrophoresis of digests
   • 08 ok
4. Ligation
   • 11(A):08(upstream)+06(downstream)+1-1G(vector)
5. Transformation of 09,10,11
6. Transfer to liquid culture:04

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