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

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Latest revision as of 18:07, 27 October 2010

E.coli Fiber Project Notebook

2010/10/23 Saturday (Naoto)

member

naoto and watachin

Experiment:Sequence

bcsA (No.9)
Big Dye
primer
DW
ethanol
EDTA
Hi-Di solution

procedure

mix materials(DNA<50ng)
PCR
Add EDTA and Ethanol and put at room temperature　(15min)
centrifuge (8000rpm,30min) and throw away supernatant
add ethanol again
centrifuge (8000rpm,15min) and throw away supernatant
add Hi-Di solution
heat at 95℃
transfer these sample to plate for sequence
read sequence

result

bcsA wasn't inserted into pSB1C3...

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

Experiment:Electrophoresis

material

PCR production (Colony PCR)

you can see other materials at protocol4

procesure

refer to protocol4

result

From the length of bands

bcsC→No.54 and 64

bcsD→No.75

seem to be correct inserts

@@ Line 1: / Line 1: @@
 {{:Team:Tokyo_Metropolitan/Notebook/Fiber}}
+__NOTOC__
 ==2010/10/23 Saturday (Naoto)==
-'''member'''
+:'''member'''
-naoto and watachin
+:naoto and watachin
+===Experiment:Sequence===
+:*bcsA (No.9)
+:*Big Dye
+:*primer
+:*DW
+:*ethanol
+:*EDTA
+:*Hi-Di solution
+:'''procedure'''
+:#mix materials(DNA<50ng)
+:#PCR
+:#Add EDTA and Ethanol and put at room temperature　(15min)
+:#centrifuge (8000rpm,30min) and throw away supernatant
+:#add ethanol again
+:#centrifuge (8000rpm,15min) and throw away supernatant
+:#add Hi-Di solution
+:#heat at 95℃
+:#transfer these sample to plate for sequence
+:#read sequence
+:'''result'''
+:bcsA wasn't inserted into pSB1C3...
 ===Experiment:Colony PCR===
-'''material'''
+:'''material'''
+:*colony of E.coli
+:**bcsB:1~32
+:**bcsC:33~64
+:**bcsD:65~96
+: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:Electrophoresis===
+:'''material'''
-*colony of E.coli
+:*PCR production (Colony PCR)
-**bcsB:1~32
+: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>
-**bcsC:33~64
-**bcsD:65~96
-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'''
+:'''procesure'''
-see <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</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'''
-From the length of bands
+:From the length of bands
-bcsC→No.54 and 64
+:bcsC→No.54 and 64
-bcsD→No.75
+:bcsD→No.75
-seem to be correct inserts
+:seem to be correct inserts
-===Preparation:making preculture===
-'''material'''
-*Colony of No.54,64,75
-*LB+Cam Broth
-'''procedure'''
-*Pick up a colony and inoculate into LB+Cam broth
+<br/>