Team:Chiba/jk/γ/week 3

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実験ログ

出力班

WEEK 3

31,August,2008

Digestion

[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]①
PCR product([http://partsregistry.org/Part:BBa_J52008 BBa_J52008])②

Sample No.	①	②
DNA	10	10
SpeⅠ	1	0
PstⅠ	1	1
XbaⅠ	0	1
BSA(x100)	0.5	0.5
Buffer2	5	0
Buffer3	0	5
dH₂O	28.5	28.5
TOTAL	50	50

-->37℃ 3hour

-->①20μl refine

-->②20μl refine

1,September,2008

Ligation

vector:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
negative control:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]②

Sample No.	①	②
vector	2	2
insert	3	0
Ligase Buffer	2	2
Ligase	1	1
dH₂O	3	5
TOTAL	10	10

-->R/T 2hour

Transformation

[http://partsregistry.org/Part:BBa_R0079 BBa_R0079](Las promoter)
[http://partsregistry.org/Part:BBa_R0071 BBa_R0071](RhlR promoter)
[http://partsregistry.org/Part:BBa_R0077 BBa_R0077](cinR promoter+RBS)
[http://partsregistry.org/Part:BBa_R0078 BBa_R0078](cinR promoter)
[http://partsregistry.org/Part:BBa_R0062 BBa_R0062](Lux promoter)

2,September,2008

TIME RESPONCE (Solid)

Colony PCR

[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]+[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]

Sample No.	①
culture	1
Fwd primer	1.5
Rev primer	1.5
Thermo pol Buffer	3
dNTP mix	3
Taq DNA pol (NEB)	0.2 (1 unit)
dH₂O	19.8
TOTAL	30

Mini prep

[http://partsregistry.org/Part:BBa_R0079 BBa_R0079]
[http://partsregistry.org/Part:BBa_R0071 BBa_R0071]
[http://partsregistry.org/Part:BBa_R0077 BBa_R0077]
[http://partsregistry.org/Part:BBa_R0078 BBa_R0078]
[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]

3,September,2008

Digestion test

[http://partsregistry.org/Part:BBa_R0079 BBa_R0079]①
[http://partsregistry.org/Part:BBa_R0071 BBa_R0071]②
[http://partsregistry.org/Part:BBa_R0077 BBa_R0077]③
[http://partsregistry.org/Part:BBa_R0078 BBa_R0078]④
[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]⑤

Sample No.	①~⑤
DNA tamplate	5
Buffer3	1
dH₂O	4
EcoRⅠ	0.1
TOTAL	10

-->37℃30min

-->Gel check

Sample No.	①~⑤
DNA(digestion sample)	10
loading Dye	2
TOTAL	12

4,Septempber,2008

Transformation

Ligation product
rLUciferase([http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] ) x3
Venus YFP([http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]) x3

5,September,2008

TIME RESPONCE(liquid)

Digestion test

[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2
[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2

DNA(digested sample)	2
PstⅠ	0.1
BSA	0.8
Buffer3	1
dH₂O	7
TOTAL	10

-->37℃ 30in

Gel Check

[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2①
[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2②
[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](PCR Product)③
[http://partsregistry.org/Part:BBa_E2050 BBa_E2050]④

Sample No.	①~②	③,④
DNA(digested sample)	10	1
loading Dye	2	1
TOTAL	12	6

Luciferase check-->no light

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