Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/19

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(Difference between revisions)

Latest revision as of 18:02, 27 October 2010

E.coli Fiber Project Notebook

2010/10/19 Tuesday (Naoto)

member

naoto and watachin

Experiment:Colony PCR

material

colony of E.coli
- bcsB:1~96
- bcsC:97~196

other materials were same as protocol3

procedure

see protocol3

Experiment:Electrophoresis

material

PCR production (Colony PCR)

you can see other materials at protocol4

procedure

refer to protocol4

result

We couldn't see bands of bcsB and bcsC...

Experiment:Sequence

material

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

procedure

mix materials(DNA<50ng)
PCR
Add EDTA and Ethanol and put at room temperature　(15min)
centrifuge (8000rpm,30min)
throw away supernatant
add ethanol again
centrifuge (8000rpm,15min)
put at refrigerator
throw away supernatant
add Hi-Di solution
transfer these sample to plate for sequence
read sequence

result

Failure to read sequence

@@ Line 2: / Line 2: @@
 __NOTOC__
 ==2010/10/19 Tuesday (Naoto)==
-'''member'''
+:'''member'''
-naoto and watachin
+:naoto and watachin
 ===Experiment:Colony PCR===
-'''material'''
+:'''material'''
-*colony of E.coli
+:*colony of E.coli
-**bcsB:1~96
+:**bcsB:1~96
-**bcsC:97~196
+:**bcsC:97~196
-other materials were same as <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
+: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'''
+:'''procedure'''
-see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
+:see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html>
 ===Experiment:Electrophoresis===
-'''material'''
+:'''material'''
-*PCR production (Colony PCR)
+:*PCR production (Colony PCR)
-you can see other materials at <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80electrophoresis">protocol4</a></html>
+:you can see other materials at <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80electrophoresis">protocol4</a></html>
-'''procedure'''
+:'''procedure'''
-refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80electrophoresis">protocol4</a></html>
+:refer to <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80electrophoresis">protocol4</a></html>
-'''result'''
+:'''result'''
-We couldn't see bands of bcsB and bcsC...
+:We couldn't see bands of bcsB and bcsC...
 ===Experiment:Sequence===
-'''material'''
+:'''material'''
-*bcsD in pSB1C3(No.89~96)
+:*bcsD in pSB1C3(No.89~96)
-*Big Dye
+:*Big Dye
-*primer
+:*primer
-*DW
+:*DW
-*ethanol
+:*ethanol
-*EDTA
+:*EDTA
-'''procedure'''
+:'''procedure'''
-#mix materials(DNA<50ng)
+:#mix materials(DNA<50ng)
-#PCR
+:#PCR
-#Add EDTA and Ethanol and put at room temperature　(15min)
+:#Add EDTA and Ethanol and put at room temperature　(15min)
-#centrifuge (8000rpm,30min)
+:#centrifuge (8000rpm,30min)
-#throw away supernatant
+:#throw away supernatant
-#add ethanol again
+:#add ethanol again
-#centrifuge (8000rpm,15min)
+:#centrifuge (8000rpm,15min)
-#put at refrigerator
+:#put at refrigerator
-#throw away supernatant
+:#throw away supernatant
-#add Hi-Di solution
+:#add Hi-Di solution
-#transfer these sample to plate for sequence
+:#transfer these sample to plate for sequence
-#read sequence
+:#read sequence
-'''result'''
+:'''result'''
-Failure to read sequence
+:Failure to read sequence
 <br/>