Team:Osaka/week13

From 2010.igem.org

October 17 (Mon)

1. To liquid culture.
   • 95, 105.
   • Pick up different colony.
2. PCR of Cel5B.
3. Ligation.
   • 082: 053, 1-23L(vector)
   • 091: 009, 1-23L(vector)
   • 094: 007, 1-23L(vector)
   • 095(C): 2-4A(upstream), 072(downstream), 1-3A(vector)
   • 105(K): 2-4A(upstream), 076(downstream), 1-5A(vector)
   • 122(A): 129(upstream), 102(downstream), 1-1C(vector)
   • 123(C): 2-4A(upstream), 102(downstream), 1-3A(vector)
   • 128: 004(upstream), 086(downstream), 1-1C(vector)
   • 129: 005(upstream), 086(downstream), 1-3A(vector)
4. Restriction digest of gel eluted Fcex1, 035 with EcoRI, SpeI.
5. Ligation
   • 079 and 039.
6. Transformation of 082(A,K),091(A,K),094(A,K),095(C),100(C),105(K),122(A),123(C),128(A),129(C).
7. Miniprep of 047, 047-2, 081, 082, 083, 092, 101, 106, 108, 036, 039.
8. Restriction digest of 047,047-2, 092, 108, 039 with EcoRI, SpeI, 081, 082, 083, 101, 036 with XbaI, PstI, 100 with EcoRI, PstI.
9. To liquid culture
   • 031-2, no-name1,2, 2-4A, BYB5013.
10. Gel Electrophoresis
    • 081 -> X
    • 082 -> OK
    • 083 -> OK
    • 092 -> X
    • 101 -> X
    • 108 -> X
    • 036 -> OK
    • 039 -> X
    • 035 -> OK
    • 051 -> X
11. Ligation
    • 085(A,K): Fcex(upstream), 1-23L(vector)
    • 092(A,K): 011, 1-23L(vector)
    • 111(C): 010(upstream), 082(downstream), 1-3A(vector)
    • 112(C): 006(upstream), 082(downstream), 1-3A(vector)
    • 113(C): 046(upstream), 082(downstream), 1-3A(vector)
    • 114(A): 004(upstream), 083(downstream), 1-1C(vector)
    • 115(A): 005(upstream), 083(downstream), 1-1C(vector)
    • 079-2(K): 079, 1-5A(vector)
    • 100①: 047(upstream), 1-23L(downstream), 1-3A(vector)
    • 100②: 039 and 079
    • 100-2(A): 039(upstream), 079(downstream), 1-1C(vector)
    • 047(A): 039(upstream), 036(downstream), 1-1C(vector)
12. PCR of Cel5B

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