Team:Osaka/week13

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)

  1. To liquid culture
  2. mini prep
    • 091①②,092①②,101①②,082①②,081①②,108①②,128①,100①①
  3. Treatment of restriction enzyme
    • 081①(EP),082①(XP),091①(EP),092①(EP),100①①(XP),101①(XP),108①(EP),128①(EP)
  4. Ligation
    • 091(C) 009 + 1-23L
    • 107(C) 2-4A + 071
    • 125(C) 010 + 1-23L
    • 132(C) 078 + 026
    • 136(C) 006 + 108.2
    • 137(C) 010 + 082
    • 031-2B(C) glr + 1-3A
    • 031-2A(C) glr + 1-3A
  5. mini prep
    • 091①②,098①②,107①②,125①②,128①②,130①②,136①②,137①②
  6. To liquid culture
    • 1-5A,1-1C,1-3Ax2
  7. Treatment of restriction enzyme
    • 091①,098①,107①,125①,128①,130①,136①,137①
  8. make culture
  9. Transformation
    • 091,107,125,132,136,137,031-2A①,031-2B①
  10. Electrophoresis
    • 081 x
    • 082 x
    • 091① ok
    • 092① x
    • 098① ok
    • 100 x
    • 101 x
    • 107
    • 108 x
    • 125 x
    • 128 ok
    • 130 x
    • 136 ok
    • 137 ok
  11. Treatment of restriction enzyme
    • 081②,092②,109②,125②,130②,094②,101②,129②,128①,108②
  12. Electrophoresis
    • PCR tube ok
    • 081 x
    • 092② ok
    • 094② ok
    • 101② ok
    • 107 x
    • 108 x
    • 125  ?
    • 128 x
    • 129 x
    • 130 x
  13. To liquid culture
    • 107③～⑤,125③～⑤,130③～⑤
  14. Treatment of restriction enzyme
    • 089①,098①,101②,131
  15. Ligation
    • 039(K) 001 + pgsB
    • 107(C) 2-4A + 071
    • 116(C) 2-4A + 102
    • 117(C) 029 + 102
    • 121(C) 078 + 099
  16. 125(C) 010 + 1-23L
  17. 129(C) 005 + 086
  18. 130 2-4A + 087

October 21 (Thu)

  1. Man26B PCR purification
  2. Treatment of restriction enzyme
    • XynA,CBM,Man,Cel5B
  3. Recovery of plasmid(mini prep)
    • 1-1C,1-3A①②,125③～⑤,130,107⑤,1-5A,107③④
  4. Transformation
    • 039,107,116,117,121,125,128,130
  5. Treatment of restriction enzyme
    • 125③④⑤,1-3A①②,1-1C,,107⑤,130③④⑤
  6. To liquid culture
    • 132,107,125,105
    • 122→x
  7. Electrophoresis
    • 130 x
    • 125 x
    • 107 x
  8. Treatment of restriction enzyme
    • 1-3A①②,1-1C, plasmid
  9. Ligation
    • 039 001 + 021
    • 100 039 + 079
    • 107(B) 2-4A + 071
    • 117 029 + 102
    • 121 078 + 099
    • 129 005 + 086
    • 130 2-4A + 087
    • 132 078 + 066
    • 129 005 + 086
    • 130 2-4A + 087
    • 125 010 + 1-23L
    • 039 001 + 021
  10. Transformation
    • 031(C)039(K),100(A),107(C),129(C)
  11. Ligation
    • Man(034),CBM(025),Cel5B(138),81(C),131,089(A)(XP)

October 22 (Fri)

  1. mini prep
    • 107①～③,107④,125①～③,132①～③,1-5A,105①～③,107③→x
  2. Transformation
    • 025(C),034(C),138(C),Y131(A),Y089(A) (昨日Ligation)
  3. Treatment of restriction enzyme
    • 132①～③,125①～③,107①～③,107④
  4. To liquid culture 17:30~
    • 039①②③,125①②③,117①②,121①②,129①,130①,116①②,BYP5013①②
  5. Electrophoresis cut check
    • 132 ok
    • 125  ?
    • 107 x

October 23 (Sat)

  1. To liquid culture
    • 131y, 189y, 025, 034, 138, 100, 129, 031, 107
  2. miniprep
    • 1-3A, 100, 039, BYB5013, 129, 125, 117, 116, 121, 125
    • 130→x
  3. Treatment of restriction enzyme
    • 125, 107
  4. Treatment of restriction enzyme
    • 1-3A,039, 100, BYB5013, 129, 125, 117
  5. cutcheck
  6. CMC plate
  7. To liquid culture
    • 034, 039, 131, 100, 089, 129, 138, 025, 030, 107, 100,
  8. ligation
    • 039 + 079
    • 130 2-4A + 087
    • 107 2-4A + 071
    • 094 1-23L + 007
    • 132 078 + 026
    • 101 075 + 1-23L
    • 129 005 + 086
    • 126 011 + 1-23L
    • 117 029 + 102
    • 123 2-4A + 102
    • 035-2 035 + 1-3A
    • 089Y 089 + BYP
    • 091-2 091 + 1-3A
    • 100 039 + 079-2
    • 132Y 132 + BYP
  9. M9 culture
  10. DNS solution


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