Team:Chiba/jk/β

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Revision as of 15:02, 11 September 2008 by Yoshimi (Talk | contribs)

β班の実験ノート

Contents

Week 1

17 August, 2008

Transformation
  • 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


Chiba-0818.JPG
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

compitent cell : 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
Chiba-0819.JPG
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
Chiba-0820-3.JPG
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


Chiba-0820-4.JPG
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



Chiba-0820.JPG
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


--->[Team:Chiba/protocol/digestion|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 12345678
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 0.10.10.10.10.10.10.10.1
PstⅠ ----0.10.10.10.1
Buffer EcoRⅠ 11111111
BSA 11111111
dH2O 4.94.94.94.94.84.84.84.8
TOTAL 1010101010101010


--->Gel Check
Chiba-0820-2.JPG
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
Chiba-0821.JPG
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
  • 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]

Compitentcell : XL10G


--->(23/8)Digestion


22 August, 2008

Transformation
  • 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
Compitentcell : XL10G


--->(24/8)Mini prep

23 August, 2008

--->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]


(21/8)--->Digestion
  • 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]


--->Gel Check</table>
Chiba-0820.JPG

From the left;
①BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
②BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
③BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
④BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK

Chiba-0823-1.JPG
  • 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]
Chiba-0823-2.JPG
  • 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]
  • M
  • 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]
Chiba-0823-3.JPG
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Single Digestion
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Double Digestion

Week 2

  • 24 August, 2008
(22/8)--->Mini prep
  • BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
  • BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
  • BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
  • BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
--->Digestion
  • BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
  • BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
  • BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
  • BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
--->Gel Check
Sample No ①②③④⑤⑥⑦⑧
Sample DNA 11111111
Loading Dye 11111111
dH2O 44444444
TOTAL 66666666
Chiba-0824-1.JPG

From the left;

⑤BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
⑥BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
⑦BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
⑧BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)

（↓single digestuion）

⑤BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
⑥BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
⑦BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
⑧BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
Sample No ⑤⑥⑦⑧
Sample DNA 9999
Loading Dye 2222
dH2O 1111
TOTAL 12121212
Chiba-0824-2.JPG

From the left; （single digestuion）

⑤BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
⑥BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
⑦BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
⑧BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
  • 25 August, 2008
(24/8)--->Digestion
①Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
②Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
③Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
④BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
Sample No ①②③④
Sample DNA 5511
EcoRⅠ 0.5o.5--
SpeⅠ 0.50.5--
Buffer 1 11--
Buffer 3 --1-
BSA 11--
dH2O 227.5-
TOTAL 1010101
--->Gel Check
Sample No ①②③④
Sample DNA 10101010
Loading Dye 2221
dH2O ---4
TOTAL 1212126
Chiba-0825-1.JPG

From right;

①Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
②Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
③Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
④BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> OK(low)


--->Digestion

①BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
②BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
③BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
]

</tr>

Sample No ①②③
Sample DNA 33ng/μl×60μl33ng/μl×60μl100ng/μl×20μl
PstⅠ 222
XbaⅠ 22-
SpeⅠ --2
Buffer 2 --5
Buffer 3 1010-
BSA 10105
dH2O 161616
TOTAL 10010052
--->Gel Check
Sample No ①②③
Sample DNA 303030
Loading Dye 666
TOTAL 363636
Chiba-0825-2.JPG
①BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
Insert:609bp -> OK
Chiba-0825-3.JPG
②BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
Insert:609bp -> OK
Chiba-0825-4.JPG
③BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
-> OK


  • 26 August, 2008

(8/25)--->Gel Extract

--->Zymo Clean
①5μl
②5μl
③27μl
--->Gel Check
Sample No ①②③
Sample DNA 222
Loading Dye 222
H2O 888
TOTAL 121212
Chiba-0825-5.JPG

From left;

①insert:C0178 -> OK
②insert:C0170 -> OK?
③Vector:J04500 -> OK


--->Dephosphorylation
③


--->Zymo Clean
③->9μl


--->Gel Check
Sample DNA ③ 1
Loading Dye 1
dH2O 4
TOTAL 6
Chiba-0826.JPG

③ -> OK ･･･30ng?


--->Ligation
]

</tr>

Sample No (1)(2)(3)(4)(5)
insert①C0178 1--1-
insert②C0170 -1--1
vector③J04500 --222
ligase 11111
Buffer 22222
dH2O 66544
TOTAL 1010101010


--->Transformation
(1)insert①C0178
(2)insert②C0170
(3)vector③J04500
(4)insert①C0178+vector③J04500
(5)insert②C0170+vector③J04500


  • 27 August, 2008

Digestion Test

Sample DNA : BBa_R0010[http://partsregistry.org/Part:BBa_R0010]
Sample No ①②③
Sample DNA 111
PstⅠ --0.5
EcoRⅠ -0.50.5
Buffer 3 -11
BSA --1
dH2O -7.52
TOTAL 11010


--->Gel Check
Sample No ①②③
Sample DNA 155
Loading Dye 111
H2O 4--
TOTAL 666
Chiba-0827.JPG

From left;

①BBa_R0010
②Single Digestion
③Double Digestion


(26/8)--->PCR
Colony PCR of 5 colonies from ligation plates (26/8-(4),(5)) and one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
]

</tr>

Sample No (4)-1~5(5)-1~5(F2620)
insert①C0178 1--
insert②C0170 -1-
vector③J04500 --2
ligase 111
Buffer 222
dH2O 665
TOTAL 101010
--->Gel Check
Sample DNA 10
Loading Dye 2
TOTAL 12
Chiba-0827-2.JPG

From left;

1~5 insert:C0170 + vector:J04500 -> OK
6 F2620･･Positive Control -> OK
7~11 insert:C0178 + vector:J04500 -> OK



--->(28/8)Mini prepDigestion Test


Transformation comptent cell : XL10G

  • BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
  • BBa_C0062(2006) [http://partsregistry.org/Part:BBa_C0062] -> no colony on plates
  • BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
  • BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
  • BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
  • BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
  • BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
  • BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
  • BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
  • BBa_S0316(2006) [http://partsregistry.org/Part:BBa_S03160]
--->Mini prep
--->(29/8)Gel Check


  • 28 August, 2008]

(27/8)--->Mini prep

①insert:C0170 + vector:J04500
②insert:C0178 + vector:J04500


--->Digestion Test
--->(29/8)Gel Check
Sample No ①②Single Digesiton:①②Double Digesiton:①②
Sample DNA 11010
Loading Dye 122
H2O 4--
TOTAL 61212
Chiba-0828.JPG

From left;

①　-> OK
②　-> OK
Single Digestion:①　-> OK
Single Digestion:②　-> OK
Double Digestion:①　-> OK
Double Digestion:②　-> OK
  • 29 August, 2008
--->Digestion Test
①BBa_R0010(2007) [http://partsregistry.org/Part:BBa_R0010]
②BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
③BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
④BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
⑤BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
⑥BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
⑦BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
⑧BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
⑨BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
⑩BBa_S03160(2006) [http://partsregistry.org/Part:BBa_S03160]


Sample No S①~⑩ｗ①ｗ②~⑩
Sample DNA 142
PstⅠ -0.0750.1
EcoRⅠ 0.10.0750.1
Buffer 3 10.751
BSA -0.751
dH2O 7.94.355.8
TOTAL 101010
--->Gel Check
Sample DNA 10
Loading Dye 2
TOTAL 12
Chiba-0829.JPG

From left;
①~⑤,S①⑤,W①~⑤

Chiba-0829-2.JPG

From left;
⑥~⑩,S⑥~⑩,W⑥~⑩


(27/8)--->

Chiba-0829-3.JPG

R0010


Digestion

①BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
②BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
③BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
④BBa_R0010(2006) [http://partsregistry.org/Part:BBa_R0010]


Sample No S①~③④
Sample DNA 100ng/μL×20200ng/μL×30
PstⅠ 22
XbaⅠ 2-
SpeⅠ -2
Buffer 2 -5
Buffer 3 5-
BSA 55
dH2O 166
TOTAL 5050
--->Gel Check
Sample DNA 103
Loading Dye 22
dH2O -7
TOTAL 12


Chiba-0829-4.JPG

From left;

Chiba-0829-5.JPG

From left;

Chiba-0829-6.JPG

From left;

Chiba-0829-7.JPG

From left;


)--->Gel Extract

--->Zymo Clean
=>1.5μL/2colum
--->SAP 
 ④
--->Zymo Clean
---> miss!


  • 30 August, 2008

Transformation comptent cell : XL10G

  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
  • BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
  • BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
  • BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
  • BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
--->(31/8)Mini prep


(29/8)--->Mini prep

①insert:C0170 + vector:J04500
②insert:C0178 + vector:J04500


--->Digestion Test
--->Gel Check


Chiba-0830.JPG

From left;

Chiba-0830-2.JPG

From left;

Chiba-0830-3.JPG

From left;

Week 3

  • 31 August, 2008
  • 1 September, 2008
  • 2 September, 2008
  • 3 September, 2008
  • 4 September, 2008
  • 5 September, 2008
  • 6 September, 2008


Week 4

  • 7 September, 2008
  • 8 September, 2008
  • 9 September, 2008
  • 10 September, 2008


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
Retrieved from "http://2008.igem.org/Team:Chiba/jk/%CE%B2"