Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/15

From 2010.igem.org

(Difference between revisions)
(New page: {{:Team:Tokyo_Metropolitan/Header}} ==2010/9/15 Wednesday (Naoto)== ===Experiment:Electrophoresis=== '''member''' naoto and watachin '''material''' *PCR production (pSB1C3) If you want...)
 
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{{:Team:Tokyo_Metropolitan/Header}}
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{{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
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__NOTOC__
==2010/9/15 Wednesday (Naoto)==
==2010/9/15 Wednesday (Naoto)==
===Experiment:Electrophoresis===
===Experiment:Electrophoresis===
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*PCR production (pSB1C3)
*PCR production (pSB1C3)
-
If you want to know other materials, see protocol4
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If you want to know other materials, see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80">the protocol4</a></html>
 +
 
'''procedure'''
'''procedure'''
-
see protocol4
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see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80Direct_PCR">the protocol4</a></html>
 +
 
'''result'''
'''result'''
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see protocol1
see protocol1
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A.xylinum(subcultured at 8/24 and 9/1)
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A.xylinum(sub-cultured at 8/24 and 9/1)
'''procedure'''
'''procedure'''
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we made 3 OWW plates and 3 tubes of broth
we made 3 OWW plates and 3 tubes of broth
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===Experiment:Purification of DNA from agarose gel with QIAGEN===
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===Experiment:Extraction of DNA from agarose gel with QIAGEN===
'''member'''
'''member'''
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*pSB1C3　in agarose gel
*pSB1C3　in agarose gel
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If you want to know other materials, see protocol4
+
If you want to know other materials, see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80">the protocol5</a></html>
 +
 
'''procedure'''
'''procedure'''
-
see protocol5
+
see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80Direct_PCR">the protocol5</a></html>
 +
 
===Experiment:PCR===
===Experiment:PCR===
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*pSB1C3,bcsB(''A.xylinum'')
*pSB1C3,bcsB(''A.xylinum'')
*a culture of ''E.coli''
*a culture of ''E.coli''
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If you want to know other materials, see protocol3(use KOD pol)
+
If you want to know other materials, see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">the protocol3</a></html>
'''procedure'''
'''procedure'''
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*add materials to PCR tubes
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#add materials to PCR tubes
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*PCR reaction (see below)
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#PCR reaction (see below)
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# nitialization :95°C 3min
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*nitialization :95°C 3min
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# denaturation:96°C 1min
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*denaturation:96°C 1min
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# annealing :51°C 1min
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*annealing :51°C 1min
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# elongation:72°C 3min
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*elongation:72°C 3min
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(30cycles from 96°C 1min to 72°C 1min)
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(30cycles from 96°C 1min to 72°C 3min)
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# reaction stop :10°C ∞
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*reaction stop :10°C ∞
===Experiment:Electrophoresis===
===Experiment:Electrophoresis===
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*PCR production (see above PCR)
*PCR production (see above PCR)
-
If you want to know other materials, see protocol4
+
If you want to know other materials, see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80Direct_PCR">the protocol4</a></html>
'''procedure'''
'''procedure'''
-
see protocol4
+
see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80Direct_PCR">the protocol4</a></html>
'''result'''
'''result'''
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#incubation at 37℃,14h
#incubation at 37℃,14h
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===Experiment:Purification of DNA from agarose gel with QIAGEN===
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===Experiment:Extraction of DNA from agarose gel===
'''member'''
'''member'''
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*pSB1C3,bcsB(''A.xylinum'') and bcsC(''E.coli'') in agarose gel
*pSB1C3,bcsB(''A.xylinum'') and bcsC(''E.coli'') in agarose gel
-
If you want to know other materials, see protocol4
+
If you want to know other materials, see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80Direct_PCR">the protocol5</a></html>
'''procedure'''
'''procedure'''
-
see protocol5
+
see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80Direct_PCR">the protocol5</a></html>
 +
 
 +
 
 +
 
 +
 
 +
<br/>

Latest revision as of 17:40, 24 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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2010/9/15 Wednesday (Naoto)

Experiment:Electrophoresis

member

naoto and watachin

material

  • PCR production (pSB1C3)

If you want to know other materials, see the protocol4


procedure

see the protocol4


result

bands could be seen slightly

Experiment:subculture of A.xylinum

member

naoto and watachin

material

see protocol1 A.xylinum(sub-cultured at 8/24 and 9/1)

procedure

see protocol1 we made 3 OWW plates and 3 tubes of broth

Experiment:Extraction of DNA from agarose gel with QIAGEN

member

naoto and watachin

material

  • pSB1C3　in agarose gel

If you want to know other materials, see the protocol5


procedure

see the protocol5


Experiment:PCR

member

naoto and watachin

material

  • pSB1C3,bcsB(A.xylinum)
  • a culture of E.coli

If you want to know other materials, see the protocol3

procedure

  1. add materials to PCR tubes
  2. PCR reaction (see below)
  • nitialization :95°C 3min
  • denaturation:96°C 1min
  • annealing :51°C 1min
  • elongation:72°C 3min

(30cycles from 96°C 1min to 72°C 3min)

  • reaction stop :10°C ∞

Experiment:Electrophoresis

member

naoto and watachin

material

  • PCR production (see above PCR)

If you want to know other materials, see the protocol4

procedure

see the protocol4

result

a band of pSB1C3 could be seen slightly bands of bcsB(A.xylinum),bcsC(E.coli) appeared clearly

Experiment:Digestion

member naoto and watachin

material

  • pSB1C3,bcsA(A.xylinum) and bcsC(E.coli)
  • 10×Mbuffer
  • BSA
  • XbaI
  • SpeI
composition
pBS1C3bcsAbcsC
solution of bcs(μl) 505050
10×Mbuffer(μl) 5 5 5
BSA(μl) 5 5 5
XbaI(μl) 1 1 1
SpeI(ul) 1 1 1

procedure

  1. add "material" to PCR tubes(show below)
  2. incubation at 37℃,14h

Experiment:Extraction of DNA from agarose gel

member

naoto and watachin

material

  • pSB1C3,bcsB(A.xylinum) and bcsC(E.coli) in agarose gel

If you want to know other materials, see the protocol5

procedure

see the protocol5




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