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From 2010.igem.org

October 17 (Sun)

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

October 18 (Mon)

1. Transformation
   • 085(A,K),092(A,B),111(C),112(C),113(C),114(A),115(A),077-2(K),100(C)①,100(C)②,100-2(A),097(A)
2. mini prep
   • 105①②,no name1,no name2,047④,2-4A,BYP5013,031-2①③④
3. Treatment of restriction enzyme
   • 047④,031-2①③④,105①②,no name1,no name2,2-4A,031-2①,BYP5013
4. Electrophoresis
   • BYP 4319bp ok
   • 105① ok
   • 105② ok
   • 047 ok
   • 031 x
   • no name2  ?
   • no name1 x
5. Treatment of restriction enzyme
   • 108②,092②,081②,101②
6. Electrophoresis
   • 108 x
   • 092 x
   • 081 x
   • 101 x
7. Ligation
   • 081(A,K) 022 + 1-23L
   • 092(A,K) 011 + 1-23L
   • 101(A,K) 075 + 1-23L
   • 108(K) 078 + 1-5A
   • Y03(C)　glr + 1-23L
8. Transformation
   • 091,128,105,122,082
9. Transformation
   • 081,092,101,108,Y03
10. culture
    • 100①～⑤,031-2⑥～⑬,039①～⑤,036,094①②,127①②,095①②,129①②,100⑥⑦+Glu,1-23L
11. Electrophoresis Cel5B
12. Ligation
    • 091(C) 009 + 1-23L
    • 107(C) 2-4A + 071
    • 098(C) 2-4A + 068
    • 125(C) 010 + 1-23L
    • 128 037 + 1-23L
    • 130 2-4A + 087
    • 136(C) 006 + 108.2
    • 137(C) 010 + 082

October 19 (Tue)

1. Transformation
   • 091,098,109,125,128,130,136,139
2. plate to liquid culture
   • 079①②,088①②,111①②,112①②,113①②,114①②,092①②,115①②
3. Recovery of plasmid (mini prep)
   • 031-2⑪,036,039①～⑤,094①②,129①②,031-2⑫,1-23L①～③
4. Treatment of restriction enzyme
   • 031-2⑪,036,039①～⑤,036
5. Electrophoresis
6. Treatment of restriction enzyme
   • 031-2⑫,1-23L①～③,BYP5013,022-2②
7. Liquid culture check 14:50~
   • 100①～⑤,031-2⑥～⑩,095①②,127①②
8. culture
   • 081①②,082①②,091①②,092①②,101①②,108①②,128①,100-2,100①②,Y03①②③
9. Treatment of restriction enzyme
   • 094①,129①,1-3A
10. Electrophoresis
    • 094① x
    • 129① x
11. Gel extraction
    • tube4 → glr
    • tube8 → XynA CBM
12. mini prep
    • 085①②,079-2①②,092①②,111①②,112①②,113①②,114①②,115①②
13. Treatment of restriction enzyme
    • 092①,111①,112①,113①,114①,115①,079-2①②,glrx2
14. Electrophoresis
    • 115 x
    • 114 x
    • 113 ok
    • 112 x
    • 111 ok
    • 092  ?
    • 079 ok

October 20 (Wed)

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