Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/20

From 2010.igem.org

(Difference between revisions)
(Experiment:Sequence)
Line 4: Line 4:
'''member'''
'''member'''
naoto and watachin
naoto and watachin
 +
 +
===Experiment:Colony PCR===
 +
'''material'''
 +
 +
*bcsB
 +
*bcsC
 +
other materials were same as <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
 +
 +
'''procedure'''
 +
 +
see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
===Experiment:Sequence===
===Experiment:Sequence===
'''material'''
'''material'''
-
*PCR
+
*bcsD in pSB1C3(No.89~96)
-
**bcsD in pSB1C3(No.89~96)
+
*Big Dye
-
**Big Dye
+
*primer
-
**primer
+
*DW
-
**DW
+
*ethanol
-
*Ethanol precipitation
+
*EDTA
-
**ethanol
+
-
**EDTA
+
'''procedure'''
'''procedure'''

Revision as of 06:58, 26 October 2010


E.coli Fiber Project Notebook

August 2010
SUNMONTUEWEDTHUFRISAT
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September 2010
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October 2010
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2010/10/20 Wednesday (Naoto)

member naoto and watachin

Experiment:Colony PCR

material

  • bcsB
  • bcsC

other materials were same as protocol3

procedure

see protocol3

Experiment:Sequence

material

  • bcsD in pSB1C3(No.89~96)
  • Big Dye
  • primer
  • DW
  • 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

result

Failure to read sequence