Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/06

From 2010.igem.org

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{{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
{{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
==2010/09/06 Monday(NEX)==
==2010/09/06 Monday(NEX)==
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===Experiment:subculture of ''E.coli''===
 
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'''Member'''<br />
 
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naoto, watachin, bambi75 and NEX
 
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'''Material'''<br />
 
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*''E.coli'' K12
 
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'''Procedure'''<br />
 
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#Pick up a culture of ''E.coli'' K12 and streak it to new culture(LB plate)
 
===Experiment:electrophoresis===
===Experiment:electrophoresis===
'''Member'''<br/>
'''Member'''<br/>
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Same above
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naoto, watachin, bambi75 and NEX
'''Material'''<br/>
'''Material'''<br/>
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'''Procedure'''<br/>
'''Procedure'''<br/>
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Follow protocol4
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refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol4</a></html>
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'''Consequence'''<br/>
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'''Result'''<br/>
Each bands were not appeared
Each bands were not appeared
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'''Procedure'''<br />
'''Procedure'''<br />
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Follow protocol3 Direct PCR
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see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
===Experiment:Transformation of pSB1C3===
===Experiment:Transformation of pSB1C3===

Revision as of 20:55, 23 October 2010


E.coli Fiber Project Notebook

August 2010
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September 2010
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October 2010
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Contents

2010/09/06 Monday(NEX)

Experiment:electrophoresis

Member
naoto, watachin, bambi75 and NEX

Material

  • PCR production (bcsA and bcsB from E.coli)
  • 1×TAE buffer
  • Agarose gel

Procedure
refer to protocol4


Result
Each bands were not appeared

Experiment:Making LB plate

Member
naoto

Material

  • Distilled water 200ml
  • LB Broth 4g

Experiment:Direct PCR

Member
naoto, watachin, bambi75 and NEX

Materials

  • sterilized water 213μl
  • Ex taq buffer 30μl
  • dNTP 24μl
  • Ex taq 3μl
  • K12bcsA sense(10μmol/l)5μl
  • K12bcsA antisense(10μmol/l)5μl
  • K12bcsB sense(10μmol/l)5μl
  • K12bcsB antisense(10μmol/l)5μl
  • K12bcsC sense(10μmol/l)5μl
  • K12bcsC antisense(10μmol/l)5μl
  • E.coli K12 strain

Procedure
see protocol3

Experiment:Transformation of pSB1C3

Member
Same above

Materials

  • pSB1C3(25ng/μl) 1μl
  • Competent cell JM109 50μl
  • LB + chloramphenicol

Procedure

  1. Mix pSB1C3 and competent cell
  2. On ice (30min)
  3. Heat shock 42℃ 45sec
  4. On ice (2min)
  5. Inoculate these onto LB plates
  6. Incubate plates at 37℃

Consequence

Failure
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