Team:Chiba/jk/β
From 2008.igem.org
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</table>
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?
[Team:Chiba/protocol/digestion|Digestion]]
- 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 ④ Sample DNA 3 10 10 Loading Dye 1 2 2 dH2O 2 - - TOTAL 6 12 12 - 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
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 ① ② ③ ④ ⑤ ⑥ ⑦ ⑧ Sample DNA 1 1 1 1 1 1 1 1 Loading Dye 1 1 1 1 1 1 1 1 dH2O 4 4 4 4 4 4 4 4 TOTAL 6 6 6 6 6 6 6 6 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 ⑤ ⑥ ⑦ ⑧ Sample DNA 9 9 9 9 Loading Dye 2 2 2 2 dH2O 1 1 1 1 TOTAL 12 12 12 12 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 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 ② ③ ④ Sample DNA 10 10 10 10 Loading Dye 2 2 2 1 dH2O - - - 4 TOTAL 12 12 12 6 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)
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 - --->Gel Check
Sample No ① ② ③ Sample DNA 30 30 30 Loading Dye 6 6 6 TOTAL 36 36 36 - ①BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
- Insert:609bp -> OK
- ①BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
- ②BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
- Insert:609bp -> OK
- ②BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
- ③BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- -> OK
- ③BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
- 26 August, 2008
(8/25)--->Gel Extract
- --->Zymo Clean
- ①5μl
- ②5μl
- ③27μl
- --->Gel Check
Sample No ① ② ③ Sample DNA 2 2 2 Loading Dye 2 2 2 H2O 8 8 8 TOTAL 12 12 12 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 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
- (1)insert①C0178
- (2)insert②C0170
- (3)vector③J04500
- (4)insert①C0178+vector③J04500
- (5)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;
- 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)).
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
- --->(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 ①② Single Digesiton:①② Double Digesiton:①② Sample DNA 1 10 10 Loading Dye 1 2 2 H2O 4 - - TOTAL 6 12 12 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 S①~⑩ 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
Sample DNA 10 Loading Dye 2 TOTAL 12 From left;
①~⑤,S①⑤,W①~⑤From left;
⑥~⑩,S⑥~⑩,W⑥~⑩
(27/8)--->R0010
- 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
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