Team:Chiba/jk/β/week 1

From 2008.igem.org

>送受信班

>Protocols

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

ホーム	メンバー紹介	プロジェクト紹介	Parts Submitted to the Registry	モデリング	ノート

Team:Chiba/jk/β/week 1

From 2008.igem.org

Contents

Week 1

17 August, 2008

18 August, 2008

19 August, 2008

20 August, 2008

21 August, 2008

22 August, 2008

23 August, 2008

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