Team:Chiba/jk/β
From 2008.igem.org
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(→27 August, 2008) |
(→27 August, 2008) |
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:(26/8)--->'''[[Team:Chiba/protocol/PCR|Colony-PCR]]''' | :(26/8)--->'''[[Team:Chiba/protocol/PCR|Colony-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)). | ::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)). | ||
- | <table width=" | + | |
+ | :<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
Line 1,294: | Line 1,295: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | |||
+ | |||
:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
Line 1,320: | Line 1,323: | ||
:--->(28/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Miniprep]]''' | :--->(28/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Miniprep]]''' | ||
- | |||
- | |||
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::*BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160] | ::*BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160] | ||
::*BBa_S0316(2006) [http://partsregistry.org/Part:BBa_S03160] | ::*BBa_S0316(2006) [http://partsregistry.org/Part:BBa_S03160] | ||
+ | |||
+ | :Compitentcell : XL10G | ||
+ | |||
:--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]''' | :--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]''' | ||
+ | |||
:--->(29/8)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | :--->(29/8)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
- | + | ||
+ | ===28 August, 2008=== | ||
:(27/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]] | :(27/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]] | ||
::#insert:C0170 + vector:J04500 | ::#insert:C0170 + vector:J04500 |
Revision as of 17:05, 11 September 2008
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
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
|
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
|
|
|
--->[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 | 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
|
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
|
- 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
- (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]
- From left; Sanple No. 1~7
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 CheckSample 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
- From left;
Week 2
24 August, 2008
- (22/8)--->Mini prep
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
- --->Digestion test
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
Sample No 5 6 7 8 BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) 1 - - - BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) - 1 - - BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) - - 1 - BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) - - - 1 EcoRⅠ 0.1 0.1 0.1 0.1 Buffer 4 0.9 0.9 0.9 0.9 dH2O 8 8 8 8 TOTAL 10 10 10 10
- --->Gel Check
Sample No 1~8 Sample DNA 1 Loading Dye 1 dH2O 4 TOTAL 6 - From left;
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> ?
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> OK
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> OK
(↓single digestuion)
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
- BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> OK
- BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> None?
- From left;
Sample No 5~8 Sample DNA 9 Loading Dye 2 dH2O 1 TOTAL 12 From the left; (single digestuion)
- 5, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
- 6, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
- 7, BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> 3400bp:Bad, 2200bp:OK
- 8, BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> OK
25 August, 2008
- (24/8)--->Digestion
- Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
- Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
- BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
Sample No 1 2 3 4 Sample DNA 5 5 1 1 EcoRⅠ 0.5 o.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:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
- Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
- Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
- BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> OK(low)
- From right;
--->Digestion (Double Digestion)- BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
- BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
- BBa_J04500[http://partsregistry.org/Part: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 </tr>
Buffer 2 - - 5 Buffer 3 10 10 - BSA 10 10 5 dH2O 16 16 16 TOTAL 100 100 52
26 August, 2008
- (8/25)--->Gel Check
Sample No 1~3 Sample DNA 30 Loading Dye 6 TOTAL 36 - 1, BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
- Insert:609bp -> OK
- 1, BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
- 2, BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
- Insert:609bp -> OK
- 2, BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
- 3, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- -> OK
- 3, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
--->Gel Extract
- --->Zymo Clean
- 5μl
- 5μl
- 27μl
- --->Gel Check
Sample No 1~3 Sample DNA 2 Loading Dye 2 H2O 8 TOTAL 12 - From left;
- insert:C0178 -> OK
- insert:C0170 -> OK?
- vector:J04500 -> OK
- From left;
- --->SAP
Sample No 3 Sample DNA 26 SAP 1 SAP Buffer 3 TOTAL 30μL
- --->Zymo Clean
- Sample No.3->9μl
- --->Gel Check
Sample No.3 1 Loading Dye 1 dH2O 4 TOTAL 6 - Sample No.3 -> OK ・・・30ng?
- --->Ligation
Sample No (1) (2) (3) (4) (5) insert①C0178 1 - - 1 - insert②C0170 - 1 - - 1 vector③J04500 - - 2 2 2 ligase 1 1 1 1 1 </tr>
Buffer 2 2 2 2 2 dH2O 6 6 5 4 4 TOTAL 10 10 10 10 10
- --->Transformation
- insert①C0178
- insert②C0170
- vector③J04500
- insert①C0178+vector③J04500
- insert②C0170+vector③J04500
27 August, 2008
- Sample DNA : BBa_R0010[http://partsregistry.org/Part:BBa_R0010]
Sample No 1 2 3 Sample DNA 1 1 1 PstⅠ - - 0.5 EcoRⅠ - 0.5 0.5 Buffer 3 - 1 1 BSA - - 1 dH2O - 7.5 2 TOTAL 1 10 10
- --->Gel Check
Sample No 1 2 3 Sample DNA 1 5 5 Loading Dye 1 1 1 H2O 4 - - TOTAL 6 6 6 - From left;
- BBa_R0010
- Single Digestion
- Double Digestion
- From left;
- (26/8)--->Colony-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)).
Sample No (4)-1~5 (5)-1~5 (F2620) insert①C0178 1 - - insert②C0170 - 1 - vector③J04500 - - 2 ligase 1 1 1 </tr>
Buffer 2 2 2 dH2O 6 6 5 TOTAL 10 10 10
- --->Gel Check
Sample DNA 10 Loading Dye 2 TOTAL 12 - From left;
- 1~5 insert:C0170 + vector:J04500 -> OK
- 6 F2620・・Positive Control -> OK
- 7~11 insert:C0178 + vector:J04500 -> OK
- From left;
- --->(28/8)Miniprep
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]
- Compitentcell : XL10G
- --->Mini prep
- --->(29/8)Gel Check
28 August, 2008
- (27/8)--->Mini prep
- insert:C0170 + vector:J04500
- insert:C0178 + vector:J04500
- --->Digestion Test
- insert:C0170 + vector:J04500
- insert:C0178 + vector:J04500
Sample No 3 4 5 6 insert:C0170 + vector:J04500 1 - 1 - insert:C0178 + vector:J04500 - 1 - 1 EcoRⅠ 0.1 0.1 0.1 0.1 SpeⅠ - - 0.1 0.1 Buffer 3 1 1 1 1 BSA - - 1 1 dH2O 7.8 7.8 6.8 6.8 TOTAL 10 10 10 10
- --->(29/8)Gel Check
Sample No 1,2 3~6 Sample DNA 1 10 Loading Dye 1 2 H2O 4 - TOTAL 6 12 - From left;
- -> OK
- -> OK
- -> OK
- -> OK
- -> OK
- -> OK
- From left;
29 August, 2008
- --->Digestion Test
- BBa_R0010(2007) [http://partsregistry.org/Part:BBa_R0010]
- BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
- 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_S03160(2006) [http://partsregistry.org/Part:BBa_S03160]
Sample No S1~10 w1 w2~10 Sample DNA 1 4 2 PstⅠ - 0.075 0.1 EcoRⅠ 0.1 0.075 0.1 Buffer 3 1 0.75 1 BSA - 0.75 1 dH2O 7.9 4.35 5.8 TOTAL 10 10 10
- --->Gel Check
width="257">Sample No. 1~10 S1~S10,W1~W10 Sample DNA 1 10 Loading Dye 1 2 TOTAL 6 12 - From left;
- 1~5,S1,S5,W1~W5
- From left;
- From left;
- 6~10,S6~S10,W6~W10
- (27/8)--->Gel Check
- <td width="257">Sample DNA</td>
<td>0.5</td> </tr> <tr> <td>Loading Dye</td> <td>1</td> </tr> <tr> <td>dH2O</td> <td>4.5</td> </tr> <tr> <td>TOTAL</td> <td>6μL</td></tr> </table>
- R0010 -> OK
- 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 S①~③ ④ Sample DNA 100ng/μL×20 200ng/μL×30 PstⅠ 2 2 XbaⅠ 2 - SpeⅠ - 2 Buffer 2 - 5 Buffer 3 5 - BSA 5 5 dH2O 16 6 TOTAL 50 50 - --->Gel Check
Sample DNA 10 3 Loading Dye 2 2 dH2O - 7 TOTAL 12
From left;
From left;
From left;
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
From left;
From left;
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
- --->Zymo Clean