Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/26

From 2010.igem.org

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{{:Team:Tokyo_Metropolitan/Header}}
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{{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
==2010/08/26(Bambi75)==
==2010/08/26(Bambi75)==
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==Make Plates==
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===Experiment:PCR===
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===member===
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'''member'''
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NEX , Bambi75 and watachin
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===Materials===
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*RO water 200ml
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*LB Broth 4g
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*Cam(50μg/L) 10ml
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===Procedure===
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① mix materials.
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② Divide ①equally and make 10 plates.
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==Separation of A.xylinum==
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===member===
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easily and naoto
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===Materials===
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*the plate(made on August 25th).
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===Procedure===
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①extract material 2ml.
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②centrifuge ① 10000rpm/5min.
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==PCR==
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===member===
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same above
same above
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===Materials===
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'''Materials'''
*2×PCR buffer  25×4μl
*2×PCR buffer  25×4μl
*2mM dNTP  10×4μl
*2mM dNTP  10×4μl
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*KOD FX 0.5×4μl
*KOD FX 0.5×4μl
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===Procedure===
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'''Procedure'''
①mix all materials for 4 tubes.
①mix all materials for 4 tubes.
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※30cycle ☆ to ★.
※30cycle ☆ to ★.
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==PCR==
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===Experiment:PCR===
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'''member'''
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===member===
 
NEX , Bambi75 and watachin
NEX , Bambi75 and watachin
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===Materials===
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'''Materials'''
*sterilized water 71μl
*sterilized water 71μl
*Ex taq buffer 10μl
*Ex taq buffer 10μl
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*primer(bcsC sense/antisense) 5μl each
*primer(bcsC sense/antisense) 5μl each
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===Procedure===
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'''Procedure'''
①mix materials
①mix materials

Revision as of 18:48, 22 October 2010


E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30 31
September 2010
SUNMONTUEWEDTHUFRISAT
1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30
October 2010
SUNMONTUEWEDTHUFRISAT
1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31

2010/08/26(Bambi75)

Experiment:PCR

member same above

Materials

  • 2×PCR buffer 25×4μl
  • 2mM dNTP 10×4μl
  • 10mM primer(sense)bcsA,B,C and D 2.5μl each
  • 10mM primer(antisense)bcsA,B,C and D 2.5μl each
  • template DNA a little
  • Q water 9×4μl
  • KOD FX 0.5×4μl

Procedure ①mix all materials for 4 tubes.

②elongation

  • bcsA,bcsB and bcsC
    • 94℃ 2min
    • 98℃ 10sec☆
    • 55℃ 30sec
    • 68℃ 4min★
    • 68℃ 7min
    • 10℃　∞
  • bcsD
    • 94℃ 2min
    • 98℃ 10sec☆
    • 55℃ 30sec
    • 68℃ 1min★
    • 68℃ 7min
    • 10℃　∞

※30cycle ☆ to ★.

Experiment:PCR

member

NEX , Bambi75 and watachin

Materials

  • sterilized water 71μl
  • Ex taq buffer 10μl
  • dNTP mix 8μl
  • Ex taq 1μl
  • primer(bcsC sense/antisense) 5μl each

Procedure ①mix materials

②divide ① into 2 tubes.

③elongation

  • 95℃ 3min
  • 96℃ 1min☆
  • 55℃ 7min
  • 72℃ 1min★
  • 10℃ ∞
※30 cycle ☆to★.
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