Team:Chiba/Calendar-Home/25 August 2008

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(Difference between revisions)

Revision as of 09:13, 24 October 2008

24 August 2008 <|> 26 August 2008

Laboratory work

Team:Input

[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](insert check済み)のグリセロールストックづくり

プレートに生えたコロニーをpickして、試験管で2ml培養(LB-Amp,37℃,12h)
培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。

OD=0.495培養液	0.67μl
60%グリセロール	0.33μl
20%グリセロールストック(計)	1.00ml

Team:Communication

(24/8)--->Digestion

Double Digestion:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)

Sample No.	1	2	3	4
Sample DNA	5	5	1	1
EcoRⅠ	0.5	0.5	-	-
SpeⅠ	0.5	0.5	-	-
Buffer 1	1	1	-	-
Buffer 3	-	-	1	-
BSA	1	1	-	-
dH₂O	2	2	7.5	-
TOTAL	10	10	10	1

--->Gel Check

Sample No.	1~3	4
Sample DNA	10	1
Loading Dye	2	1
dH₂O	-	4
TOTAL	12	6

From right;

Double Digestion:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007) -> OK
Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> OK(low)

--->Digestion (Double Digestion)

[http://partsregistry.org/Part:BBa_C0178 BBa_C0178]
[http://partsregistry.org/Part:BBa_C0170 BBa_C0170]
[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)

Sample No.	1	2	3
Sample DNA	33ng/μl×60μl	33ng/μl×60μl	100ng/μl×20μl
PstⅠ	2	2	2
XbaⅠ	2	2	-
SpeⅠ	-	-	2
Buffer 2	-	-	5
Buffer 3	10	10	-
BSA	10	10	5
dH₂O	16	16	16
TOTAL	100	100	52

Team:Output

PCR

[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
BL21③
[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④
[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤

>

Sample No.	①	②	③	④	⑤
DNA tamplate	1	1	1	1	1
FW primer	5(Venus_YFP_fwd)	5(LacZ_fwd)	5(LacZ_fwd)	5(rLUC_fwd)	5(fLuc_fwd)
VR primer	5(VR)	5(LacZ_rev)	5(LacZ_rev)	5(VR)	5(VR)
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
DNA tamplate	1	1	1	1	1
loading Dye	1	1	1	1	1
dH₂O	4	4	4	4	4
TOTAL	6	6	6	6	6

Mini prep

Venus
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]

Transformation

Venus
PUC19
pGFPUV
pGEX Venus YFP
Membren Venus YFP

PCR

[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]①

Sample No.	①
DNA tamplate	1
FW primer(Venus)	5
VR primer	5
dNTPmix	10
Thermo pol buffer	10
DNA pol(VENT)	1
dH₂O	68
TOTAL	100

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 ==Laboratory work==