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

From 2010.igem.org

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==2010/09/06 Monday(NEX)==
==2010/09/06 Monday(NEX)==
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===Experiment:electrophoresis===
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===Experiment:Electrophoresis===
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'''Member'''<br/>
+
:'''Member'''<br/>
-
naoto, watachin, bambi75 and NEX
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:naoto, watachin, bambi75 and NEX
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'''Material'''<br/>
+
:'''Material'''<br/>
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*PCR production (bcsA and bcsB from E.coli)
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:*PCR production (bcsA and bcsB)
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*1×TAE buffer
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:*1×TAE buffer
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*Agarose gel
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:*Agarose gel
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'''Procedure'''<br/>
+
:'''Procedure'''<br/>
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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>  
+
:refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol4</a></html>  
-
'''Result'''<br/>
+
:'''Result'''<br/>
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Each bands were not appeared
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:Each bands were not appeared
 +
===Preparation:Making LB plate===
 +
:'''Member'''<br />
 +
:naoto
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===Experiment:Making LB plate===
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:'''Material'''
-
'''Member'''<br />
+
-
naoto
+
-
'''Material'''
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:see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol1</a></html>
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*Distilled water 200ml
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*LB Broth 4g
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 +
:'''Procedure'''
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===Experiment:Direct PCR===
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:refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol1</a></html>  
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'''Member'''<br />
+
-
naoto, watachin, bambi75 and NEX
+
-
'''Materials'''<br />
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===Experiment:PCR===
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*sterilized water 213μl
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:'''Member'''<br />
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*Ex taq buffer 30μl
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:naoto, watachin, bambi75 and NEX
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*dNTP 24μl
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-
*Ex taq 3μl
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-
*K12bcsA sense(10μmol/l)5μl
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*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'''<br />
+
:'''Material'''<br />
-
see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
+
:*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'''<br />
 +
: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===
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'''Member'''<br />
+
:'''Member'''<br />
-
Same above
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:Same above
-
 
+
-
'''Materials'''<br />
+
-
*pSB1C3(25ng/μl) 1μl
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-
*Competent cell JM109 50μl
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*LB + chloramphenicol
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-
 
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'''Procedure'''<br />
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#Mix pSB1C3 and competent cell
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#On ice (30min)
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#Heat shock 42℃ 45sec
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#On ice (2min)
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#Inoculate these onto LB plates
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-
#Incubate plates at 37℃
+
-
 
+
-
'''Consequence'''<br />
+
-
Failure
+
 +
:'''Materials'''<br />
 +
:*pSB1C3(25ng/μl) 1μl
 +
:*Competent cell JM109 50μl
 +
:*LB + Chloramphenicol
 +
:'''Procedure'''<br />
 +
:#Mix pSB1C3 and competent cell
 +
:#On ice (30min)
 +
:#Heat shock 42℃ 45sec
 +
:#On ice (2min)
 +
:#Inoculate onto LB plates
 +
:#Incubate plates at 37℃
 +
:'''Consequence'''<br />
 +
:Failure
<br/>
<br/>

Latest revision as of 06:34, 27 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

2010/09/06 Monday(NEX)

Experiment:Electrophoresis

Member
naoto, watachin, bambi75 and NEX
Material
  • PCR production (bcsA and bcsB)
  • 1×TAE buffer
  • Agarose gel
Procedure
refer to protocol4
Result
Each bands were not appeared

Preparation:Making LB plate

Member
naoto
Material
see protocol1
Procedure
refer to protocol1

Experiment:PCR

Member
naoto, watachin, bambi75 and NEX
Material
  • 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 onto LB plates
  6. Incubate plates at 37℃
Consequence
Failure

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