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	>[[Team:Chiba/jk/β|送受信班]]		>[[Team:Chiba/jk/β|送受信班]]
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		+	>[[Team:Chiba/protocol/j|Protocols]]
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	==Week 1==		==Week 1==

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Revision as of 07:46, 30 September 2008

>送受信班

>Protocols

Contents 1 Week 1 1.1 17 August, 2008 1.2 18 August, 2008 1.3 19 August, 2008 1.4 20 August, 2008 1.5 21 August, 2008 1.6 22 August, 2008 1.7 23 August, 2008 1.8 >next week

Week 1

17 August, 2008

Transformation

competent cells : XL10G

• BBa_R0040(2008) [http://partsregistry.org/Part:BBa_R0040] -> No colonies on plates
• BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070] -> No colonies on plates
• BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
• BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
• BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
• BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079] -> No colonies on plates

18 August, 2008

PCR

• BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070]
• BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076]
• BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077]
• BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078]
• BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079]
• BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]･･･control

DNA Template	1
dNTP mix	10
Foward Primer	5
Reverse Primer	5
DNA polymerase	1
Thermopol Buffer	10
dH2O	68
TOTAL	100μL

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070] ->None BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] ->None BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] ->None BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] ->None BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079] ->None BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]･･･control ->OK

Transformation

competent cells : XL10G

• BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
• BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
• BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
• BBa_C0076(2006) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates(19/8)
• BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
• BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
• BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
• BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
• BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
• BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]

--->(20/8)Mini prep

19 August, 2008

PCR

• BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
• BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
• BBa_S3154(2007) [http://partsregistry.org/Part:BBa_S3154]
• BBa_S3154(2006) [http://partsregistry.org/Part:BBa_S3154]
• BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]･･･control

DNA Template	1
dNTP mix	10
Foward Primer	5
Reverse Primer	5
DNA polymerase VENT	1
Thermopol Buffer	5
dH2O	28
TOTAL	50μL

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] ->None BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] ->None BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] ->None BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] ->None BBa_S3154(2007) [http://partsregistry.org/Part:BBa_S3154] ->None BBa_S3154(2006) [http://partsregistry.org/Part:BBa_S3154] ->None BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]･･･control ->OK

20 August, 2008

(18/8)--->Mini prep

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

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None

Sample DNA 10 Loading Dye 4 dH2O 10 TOTAL 24μL From left; BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> OK BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK

--->Digestion test

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

Sample No	1	2	3	4	5	6	7	8
BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]	3	-	-	-	3	-	-	-
BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]	-	3	-	-	-	3	-	-
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]	-	-	3	-	-	-	3	-
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]	-	-	-	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) BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK (Double Digestion 1~4) BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK

21 August, 2008

(20/8)--->PCR

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

DNA Template	1
dNTP mix	10
Foward Primer	5
Reverse Primer	5
DNA polymerase TAQ	1
Thermopol Buffer	5
dH2O	28
TOTAL	50μL

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

--->Gel Check

Sample DNA 3 Loading Dye 2 dH2O 7 TOTAL 12μL From left; BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK

Transformation

competent cells : XL10G

• BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
• BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
• BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
• BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]

--->(23/8)Digestion

22 August, 2008

Transformation

competent cells : XL10G

• BBa_J13002(2007) [http://partsregistry.org/Part:BBa_J13002]
• BBa_J13002(2006) [http://partsregistry.org/Part:BBa_J13002]
• BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
• BBa_J04500(2006) [http://partsregistry.org/Part:BBa_J04500]

(Locate & 2 Punch ~iyama ver.~)

• BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
• BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
• BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
• BBa_J04500(2008) [http://partsregistry.org/Part:BBa_J04500] -> No colonies on plates

--->(24/8)Mini prep

23 August, 2008

(21/8)--->Mini prep

• BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
• BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
• BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
• BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]

--->Digestion test

• BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
• BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
• BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
• BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
• BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
• BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]

Sample No	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-
BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-
BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-
BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-
BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-
BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]	-	-	-	-	-	3	-	-	-	-	-	-	3	-	-	-	-	-	-	3	-
BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]	-	-	-	-	-	-	1	-	-	-	-	-	-	3	-	-	-	-	-	-	3
EcoRⅠ	-	-	-	-	-	-	-	0.1	0.1	0.1	0.1	0.1	0.1	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	0.1	0.1	0.1
Buffer EcoRⅠ	-	-	-	-	-	-	-	0.9	0.9	0.9	0.9	0.9	0.9	0.9	0.8	0.8	0.8	0.8	0.8	0.8	0.8
BSA	-	-	-	-	-	-	-	1	1	1	1	1	1	1	1	1	1	1	1	1	1
dH2O	-	-	-	-	-	-	-	5	5	5	5	5	5	5	5	5	5	5	5	5	5
TOTAL	3	3	3	3	3	3	1	10	10	10	10	10	10	10	10	10	10	10	10	10	10

--->Gel Check

Sample No 1~6 7 Sample DNA 3 1 Loading Dye 1 1 dH2O 2 4 TOTAL 6 6 From left; Sanple No. 1~7 BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]

Sample No 8~21 Sample DNA 10 Loading Dye 2 TOTAL 12 From left; (Single Digestion) Sanple No. 8~14 BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] -> OK BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] -> OK BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] -> OK BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] -> OK BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] -> OK BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] -> OK BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK (Double Digestion) Sample No. 15~21 BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] -> OK BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] -> OK BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] -> OK BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] -> OK BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] -> OK BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] -> OK BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK?

Digestion test

BBa_J04500(2007:old-sample)[http://partsregistry.org/Part:BBa_J04500]

Sample No	1	2	3
Sample DNA	3	3	3
EcoRⅠ	-	0.1	0.1
SpeⅠ	-	-	0.1
EcoRⅠ Buffer	-	0.9	0.8
BSA	-	1	1
dH2O	-	5	5
TOTAL	3	10	10

--->Gel Check

Sample No 1 2 3 Sample DNA 3 10 10 Loading Dye 1 2 2 dH2O 2 - - TOTAL 6 12 12 From left; BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Single Digestion -> OK BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Double Digestion -> Bad

>next week

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