Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/01

From 2010.igem.org

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{{:Team:Tokyo_Metropolitan/Header}}
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{{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
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==2010/9/1 Wednesday(Naoto)==
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__NOTOC__
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==2010/09/01 Wednesday(Naoto)==
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===Experiment:digestion of bcsA,B,C,D(''A.xylinum'')===
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===Experiment:Digestion of bcsA,B,C and D===
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'''member'''
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:'''Member'''
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naoto
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:naoto
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'''material'''
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:'''Material'''
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*bcsA,B,C,D(from ''A.xylinum'')
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:*bcsA,B,C,D
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*10×Mbuffer
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:*10×Mbuffer
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*BSA
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:*BSA
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*XbaI
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:*XbaI
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*SpeI
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:*SpeI
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'''procedure'''
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:'''Procedure'''
#add "material" to PCR tubes(show below)
#add "material" to PCR tubes(show below)
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===Preparation:Subculture of''A.xylinum''===
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:'''Member'''
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===Experiment:subculture of''A.xylinum''===
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:naoto
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'''member'''
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naoto
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:'''Material'''
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'''material'''
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:''A.xylinum'' JCM7664
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''A.xylinum'' JCM7664
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:'''Procedure'''
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'''procedure'''
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:transfer ''A.xylinum'' JCM7664 to new culture(OWW and Broth 8/25 made)
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transfer ''A.xylinum'' JCM7664 to new culture(OWW and JCM Broth 8/25 made)
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===Experiment:PCR===
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:'''Member'''
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:naoto
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===Experiment:subculture of ''E.coli''===
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:'''Material'''
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'''member'''
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naoto
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:* ''E.coli'' K12 strain
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:* 2×PCR buffer 25μl×2
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:* 2mM dNTP 10μl×2
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:* 10μM primer(sense)bcsA,B 2.5μl each
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:* 10μM primer(antisense)bcsA,B 2.5μl each
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:* milli-Q water 9μl×2
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:* KOD FX 0.5μl×2
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'''material'''
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:'''Procedure'''
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''E.coli'' K12
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:follow <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
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'''procedure'''
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<br/>
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pick up a culture of ''E.coli'' K12 and streak it to new culture(LB plate)
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===Experiment:direct PCR===
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'''member'''
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naoto
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'''material'''
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* ''E.coli'' K12
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* 2×PCR buffer 25μl×2
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* 2mM dNTP 10μl×2
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* 10μM primer(sense)bcsA,B 2.5μl each
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* 10μM primer(antisense)bcsA,B 2.5μl each
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* milli-Q water 9μl×2
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* KOD FX 0.5μl×2
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'''procedure'''
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follow protocol3 direct PCR
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Latest revision as of 22:02, 26 October 2010


E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
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29 30 31
September 2010
SUNMONTUEWEDTHUFRISAT
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12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30
October 2010
SUNMONTUEWEDTHUFRISAT
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10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31

2010/09/01 Wednesday(Naoto)

Experiment:Digestion of bcsA,B,C and D

Member
naoto
Material
  • bcsA,B,C,D
  • 10×Mbuffer
  • BSA
  • XbaI
  • SpeI
Procedure
  1. add "material" to PCR tubes(show below)
  2. incubation at (37℃,7h)
composition
bcsAbcsBbcsCbcsD
solution of bcs(μl) 20202020
10×Mbuffer(μl) 2 2 2 2
BSA(μl) 2 2
XbaI(μl) 0.8 0.8
SpeI(ul) 0.8 0.8

Preparation:Subculture ofA.xylinum

Member
naoto
Material
A.xylinum JCM7664
Procedure
transfer A.xylinum JCM7664 to new culture(OWW and Broth 8/25 made)

Experiment:PCR

Member
naoto
Material
  • E.coli K12 strain
  • 2×PCR buffer 25μl×2
  • 2mM dNTP 10μl×2
  • 10μM primer(sense)bcsA,B 2.5μl each
  • 10μM primer(antisense)bcsA,B 2.5μl each
  • milli-Q water 9μl×2
  • KOD FX 0.5μl×2
Procedure
follow protocol3

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