Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/23

From 2010.igem.org

(Difference between revisions)
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naoto and watachin
naoto and watachin
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===Experiment:Sequence===
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'''material'''
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For PCR
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*bcsA (No.9)
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*Big Dye
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*primer
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*DW
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For Ethanol precipitation
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*ethanol
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*EDTA
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'''procedure'''
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#mix materials(DNA<50ng)
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#PCR
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#Add EDTA and Ethanol and put at room temperature　(15min)
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#centrifuge (8000rpm,30min)
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#throw away supernatant
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#add ethanol again
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#centrifuge (8000rpm,15min)
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#put at refrigerator
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#throw away supernatant
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#add Hi-Di solution
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#transfer these sample to plate for sequence
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#read sequence
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<br/>
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===Experiment:Colony PCR===
===Experiment:Colony PCR===
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seem to be correct inserts
seem to be correct inserts
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===Preparation:making preculture===
 
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'''material'''
 
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*Colony of No.54,64,75
 
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*LB+Cam Broth
 
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'''procedure'''
 
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*Pick up a colony and inoculate into LB+Cam broth
 
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<br/>
<br/>

Revision as of 16:39, 25 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
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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

2010/10/23 Saturday (Naoto)

member

naoto and watachin

Experiment:Sequence

material

For PCR

  • bcsA (No.9)
  • Big Dye
  • primer
  • DW

For Ethanol precipitation

  • ethanol
  • EDTA

procedure

  1. mix materials(DNA<50ng)
  2. PCR
  3. Add EDTA and Ethanol and put at room temperature　(15min)
  4. centrifuge (8000rpm,30min)
  5. throw away supernatant
  6. add ethanol again
  7. centrifuge (8000rpm,15min)
  8. put at refrigerator
  9. throw away supernatant
  10. add Hi-Di solution
  11. transfer these sample to plate for sequence
  12. read sequence








Experiment:Colony PCR

material

  • colony of E.coli
    • bcsB:1~32
    • bcsC:33~64
    • bcsD:65~96

other materials were same as protocol3

procedure

see protocol3

result

From the length of bands

bcsC→No.54 and 64

bcsD→No.75

seem to be correct inserts



Retrieved from "http://2010.igem.org/Team:Tokyo_Metropolitan/Notebook/Fiber/2010/10/23"