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

From 2010.igem.org

(Difference between revisions)
(New page: ==2010/09/06 Monday(NEX)== ===Experiment:subculture of ''E.coli''=== '''member'''<br /> naoto, watachin, bambi75 and NEX '''material'''<br /> *''E.coli'' K12 '''procedure'''<br /> #Pick ...)
(→Experiment:electrophoresis)
 
(12 intermediate revisions not shown)
Line 1: Line 1:
 +
{{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
 +
__NOTOC__
==2010/09/06 Monday(NEX)==
==2010/09/06 Monday(NEX)==
-
===Experiment:subculture of ''E.coli''===
 
-
'''member'''<br />
 
-
naoto, watachin, bambi75 and NEX
 
-
'''material'''<br />
+
===Experiment:Electrophoresis===
-
*''E.coli'' K12
+
:'''Member'''<br/>
 +
:naoto, watachin, bambi75 and NEX
-
'''procedure'''<br />
+
:'''Material'''<br/>
-
#Pick up a culture of ''E.coli'' K12 and streak it to new culture(LB plate)
+
:*PCR production (bcsA and bcsB)
 +
:*1×TAE buffer
 +
:*Agarose gel
-
===Experiment:electrophoresis===
+
:'''Procedure'''<br/>
-
'''member'''<br/>
+
:refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol4</a></html>
-
Same above
+
-
'''material'''<br/>
+
:'''Result'''<br/>
-
*PCR production (bcsA and bcsB from E.coli)
+
:Each bands were not appeared
-
*1×TAE buffer
+
-
*Agarose gel
+
-
'''procedure'''<br/>
+
===Preparation:Making LB plate===
-
Follow to protocol4
+
:'''Member'''<br />
 +
:naoto
-
'''result'''<br/>
+
:'''Material'''
-
Each bands were not appeared
+
-
===Experiment:Making LB plate===
+
:see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol1</a></html>  
-
'''Member'''<br />
+
-
naoto
+
-
'''Material'''
+
:'''Procedure'''
-
*Distilled water 200ml
+
-
*LB Broth 4g
+
-
===Experiment===:Direct PCR===
+
:refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol1</a></html>  
-
'''Member'''<br />
+
-
naoto, watachin, bambi75 and NEX
+
-
'''Materials'''<br />
+
===Experiment:PCR===
-
*sterilized water 213μl
+
:'''Member'''<br />
-
*Ex taq buffer 30μl
+
:naoto, watachin, bambi75 and NEX
-
*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 />
+
:'''Material'''<br />
-
Follow to protocol3 Direct PCR
+
:*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
-
===Experiment===
+
:'''Procedure'''<br />
-
'''Member'''<br />
+
:see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
-
Same above
+
-
'''Materials'''<br />
 
-
*pSB1A3(25ng/μl) 1μl
 
-
*Competent cell JM109 50μl
 
-
*LB + chloramphenicol
 
-
'''Procedure'''<br />
+
===Experiment:Transformation of pSB1C3===
-
#Mix pSB1A3 and competent cell
+
:'''Member'''<br />
-
#On ice (30min)
+
:Same above
-
#Heat shock 42℃ 45sec
+
-
#On ice (2min)
+
-
#Inoculate these onto LB plates
+
-
#Incubate plates at 37℃
+
-
'''Consequence'''<br />
+
:'''Materials'''<br />
-
Failure
+
:*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/>

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

Retrieved from "http://2010.igem.org/Team:Tokyo_Metropolitan/Notebook/Fiber/2010/09/06"