Team:Chiba/Calendar-Home/20 August 2008

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Laboratory work

Team:Input

PCR

[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)①
[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2008)②
[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)③
[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2008)④
[http://partsregistry.org/Part:BBa_E0612 BBa_E0612](2008)⑤

>

Sample No.	①	②	③	④	⑤
DNA tamplate	1	1	1	1	1
FW primer	5	5	5	5	5
VR primer	5	5	5	5	5
dNTPmix	10	10	10	10	10
thermo pol buffer	10	10	10	10	10
DNA pol(VENT)	1	1	1	1	1
dH₂O	68	68	68	68	68
TOTAL	100	100	100	100	100

-->Gel Check

>

Sample No.	①	②	③	④	⑤
Sample DNA	1	1	1	1	1
loading dye	1	1	1	1	1
dH₂O	4	4	4	4	4
Total	6	6	6	6	6

-->どれもバンド出ず。

Transformation

[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)

-->BBa_J22141のみコロニーがほとんど生えていなかった。(１個のみ)

Team:Communication

(18/8)--->Mini prep

[http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2007)
[http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2006)
[http://partsregistry.org/Part:BBa_C0076 BBa_C0076](2007)
[http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2007)
[http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2006)
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2007)
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2006)
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2007)
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2006)

--->Gel Check

Sample DNA	3
Loading Dye	2
dH₂O	7
TOTAL	12μL

From left;

[http://partsregistry.org/Part:BBa_C0076 BBa_C0076](2007) -> None
[http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2007) -> None
[http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2006) -> None

Sample DNA	10
Loading Dye	4
dH₂O	10
TOTAL	24μL

From left;

[http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2007) -> None
[http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2006) -> OK
[http://partsregistry.org/Part:BBa_C0076 BBa_C0076](2007) -> None
[http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2007) -> OK
[http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2006) -> OK

Sample DNA	3
Loading Dye	2
dH₂O	7
TOTAL	12μL

From left;

[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2007) -> OK
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2006) -> OK
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2007) -> OK
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2006) -> OK

--->Digestion test

[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2007)
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2006)
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2007)
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2006)

Sample No	1	2	3	4	5	6	7	8
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2006)	3	-	-	-	3	-	-	-
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2007)	-	3	-	-	-	3	-	-
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2006)	-	-	3	-	-	-	3	-
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2007)	-	-	-	3	-	-	-	3
EcoRⅠ	0.1	0.1	0.1	0.1	0.1	0.1	0.1	0.1
PstⅠ	-	-	-	-	0.1	0.1	0.1	0.1
Buffer EcoRⅠ	1	1	1	1	1	1	1	1
BSA	1	1	1	1	1	1	1	1
dH₂O	4.9	4.9	4.9	4.9	4.8	4.8	4.8	4.8
TOTAL	10	10	10	10	10	10	10	10

--->Gel Check

Sample DNA	3
Loading Dye	2
dH₂O	7
TOTAL	12μL

From left;

(Single Digestion 1~4)

[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2006) -> OK
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2007) -> OK
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2006) -> OK
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2007) -> OK

(Double Digestion 1~4)

[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2006) -> OK
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](2007) -> OK
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2006) -> OK
[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](2007) -> OK

Team:Output

Transformation

[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](Luciferase Renilla)-->colony
[http://partsregistry.org/Part:BBa_I712019 BBa_I712019](Luciferase Fire fly)-->no colony
[http://partsregistry.org/Part:BBa_J33202 BBa_J33202](LacZ)-->colony
[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](Venus YFP)-->not so good
YFP(P-GEX)-->colony
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]-->colony
[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]-->colony

Luciferase(J52008:2007,2008 I712019:2008)(Amp)
Lac Z(J33202:2007,2008)(Amp)
Venus(E2030:2007,2008)(Kam)
Craig Venus(Amp?Kan?)
T9002(Amp)
F2620(Amp)

luciferase,LacZ,Venus
competent cell 50μl
DNA 1μl
soc 200μl
plating 250μl

Craig Venus,T9002,F2620
competent cell 30μl
DNA 1μl
soc 120μl
plating 20μl

result

成功→Luciferase(J52008:2007),Lac Z(J33202:2007),Craig Venus,T9002,F2620

失敗→Luciferase(J52008:2008 J52008:2008),Lac Z(J33202:2008),Venus(J32007:2007,2008)

Team:Chiba/Calendar-Home/20 August 2008

From 2008.igem.org

Contents

Laboratory work

Team:Input

Team:Communication

Team:Output