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From 2008.igem.org

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27 August 2008 <|> 29 August 2008

Contents 1 Laboratory work 1.1 Team:Input 1.2 Team:Communication 1.3 Team:Output

Laboratory work

Team:Input

Transformation

[http://partsregistry.org/Part:BBa_I7106 BBa_I7106](2007) 生えてきたコロニーを10ml培養してMini prep(60mlで溶出)

-->Digestion
混ぜ表

	double
dH2O	2
10×BSA	1
10×NE	1
EcoRI	0.5
PstI	0.5
DNA	5
TOTAL	10μl

Digestion 37°C、30分
プラスミドの濃度は33.6ng/μl

PCR
RBS＋CI
混ぜ表

DNA	1
FW primer	2.5
VR primer	2.5
dNTPmix	5
thermo pol buffer	5
DNA pol(VENT)	1
dH2O	34
TOTAL	50μl

Gel Check

PCR産物	1
Dye	1
dH2O	4
TOTAL	6μl

-->Concentration:33.6ng/μl

VectorPCR
[http://partsregistry.org/Part:BBa_I7106 BBa_I7106]

DNA	1
FW primer	5
VR primer	5
dNTPmix	10
thermo pol buffer	10
DNA pol(VENT)	1
dH2O	68
TOTAL	100μl

Gel check

PCR産物	1
Dye	1
dH2O	4
TOTAL	6μl

-->バンド出ず。

Team:Communication

(27/8)--->Mini prep

1. insert:[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
2. insert:[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]

--->Digestion Test

1. insert:[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
2. insert:[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] + vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]

Sample No.	3	4	5	6
insert:C0170 + vector:J04500	1	-	1	-
insert:C0178 + vector:J04500	-	1	-	1
EcoRⅠ	0.1	0.1	0.1	0.1
SpeⅠ	-	-	0.1	0.1
Buffer 3	1	1	1	1
BSA	-	-	1	1
dH2O	7.8	7.8	6.8	6.8
TOTAL	10	10	10	10

--->(29/8)Gel Check

Sample No. 1,2 3~6 Sample DNA 1 10 Loading Dye 1 2 H2O 4 - TOTAL 6 12 From left; 　-> OK 　-> OK 　-> OK 　-> OK 　-> OK 　-> OK

Team:Output

Time Responce(liquid)

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