Team:Osaka/week13

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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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