Team:Chiba/jk/γ/week 3
From 2008.igem.org
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[[Team:Chiba/jk|実験ログ]] | [[Team:Chiba/jk|実験ログ]] | ||
- | + | ||
+ | [[Team:Chiba/jk/γ|出力班]] | ||
== WEEK 3 == | == WEEK 3 == | ||
Line 9: | Line 9: | ||
== 31,August,2008 == | == 31,August,2008 == | ||
- | + | [[Team:Chiba/protocol/digestion|Digestion]] | |
- | + | ||
- | + | *[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]① | |
+ | *PCR product([http://partsregistry.org/Part:BBa_J52008 BBa_J52008])② | ||
+ | |||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">Sample No</td> | + | <td width="257">Sample No.</td> |
<td>①</td><td>②</td> | <td>①</td><td>②</td> | ||
</tr> | </tr> | ||
Line 46: | Line 48: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>28.5</td><td>28.5</td> | <td>28.5</td><td>28.5</td> | ||
</tr> | </tr> | ||
Line 54: | Line 56: | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
- | + | -->37℃ 3hour | |
- | + | ||
+ | -->①20μl refine | ||
+ | |||
+ | -->②20μl refine | ||
== 1,September,2008 == | == 1,September,2008 == | ||
- | + | [[Team:Chiba/protocol/ligation/ligation|Ligation]] | |
- | + | *vector:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]① | |
- | + | *negative control:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]② | |
+ | |||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">Sample No</td> | + | <td width="257">Sample No.</td> |
<td>①</td><td>②</td> | <td>①</td><td>②</td> | ||
</tr> | </tr> | ||
Line 86: | Line 92: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>3</td><td>5</td> | <td>3</td><td>5</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
<td>10</td><td>10</td> | <td>10</td><td>10</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | |||
- | + | -->R/T 2hour | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
+ | [[Team:Chiba/protocol/transformation|Transformation]] | ||
+ | |||
+ | *[http://partsregistry.org/Part:BBa_R0079 BBa_R0079](Las promoter) | ||
+ | *[http://partsregistry.org/Part:BBa_R0071 BBa_R0071](RhlR promoter) | ||
+ | *[http://partsregistry.org/Part:BBa_R0077 BBa_R0077](cinR promoter+RBS) | ||
+ | *[http://partsregistry.org/Part:BBa_R0078 BBa_R0078](cinR promoter) | ||
+ | *[http://partsregistry.org/Part:BBa_R0062 BBa_R0062](Lux promoter) | ||
Line 107: | Line 115: | ||
== 2,September,2008 == | == 2,September,2008 == | ||
- | + | [[Team:Chiba/jk/γ/trs|TIME RESPONCE (Solid)]] | |
+ | |||
+ | Colony PCR | ||
+ | *[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]+[http://partsregistry.org/Part:BBa_J52008 BBa_J52008] | ||
- | |||
- | |||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">Sample No</td> | + | <td width="257">Sample No.</td> |
<td>①</td> | <td>①</td> | ||
</tr> | </tr> | ||
Line 120: | Line 129: | ||
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
<td>Fwd primer</td> | <td>Fwd primer</td> | ||
<td>1.5</td> | <td>1.5</td> | ||
Line 140: | Line 150: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>19.8</td> | <td>19.8</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
<td>30</td> | <td>30</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
- | + | [[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]] | |
- | + | *[http://partsregistry.org/Part:BBa_R0079 BBa_R0079] | |
- | + | *[http://partsregistry.org/Part:BBa_R0071 BBa_R0071] | |
- | + | *[http://partsregistry.org/Part:BBa_R0077 BBa_R0077] | |
- | + | *[http://partsregistry.org/Part:BBa_R0078 BBa_R0078] | |
+ | *[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] | ||
== 3,September,2008 == | == 3,September,2008 == | ||
- | + | [[Team:Chiba/protocol/digestion|Digestion test]] | |
- | + | *[http://partsregistry.org/Part:BBa_R0079 BBa_R0079]① | |
- | + | *[http://partsregistry.org/Part:BBa_R0071 BBa_R0071]② | |
- | + | *[http://partsregistry.org/Part:BBa_R0077 BBa_R0077]③ | |
- | + | *[http://partsregistry.org/Part:BBa_R0078 BBa_R0078]④ | |
- | + | *[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]⑤ | |
+ | |||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">Sample No</td> | + | <td width="257">Sample No.</td> |
<td>①~⑤</td> | <td>①~⑤</td> | ||
</tr> | </tr> | ||
Line 176: | Line 189: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>4</td> | <td>4</td> | ||
</tr> | </tr> | ||
Line 183: | Line 196: | ||
<td>0.1</td> | <td>0.1</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
<td>10</td> | <td>10</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
- | + | -->37℃30min | |
+ | |||
+ | -->[[Team:Chiba/protocol/gelcheck|Gel check]] | ||
+ | |||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">Sample No</td> | + | <td width="257">Sample No.</td> |
<td>①~⑤</td> | <td>①~⑤</td> | ||
</tr> | </tr> | ||
Line 208: | Line 225: | ||
</table> | </table> | ||
+ | == 4,Septempber,2008 == | ||
+ | [[Team:Chiba/protocol/transformation|Transformation]] | ||
- | + | *Ligation product | |
- | + | *rLUciferase([http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] ) x3 | |
- | + | *Venus YFP([http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]) x3 | |
- | + | ||
- | + | ||
== 5,September,2008 == | == 5,September,2008 == | ||
- | + | [[Team:Chiba/jk/γ/Trl|TIME RESPONCE(liquid)]] | |
- | + | ||
- | + | [[Team:Chiba/protocol/digestion|Digestion test]] | |
- | + | *[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2 | |
+ | *[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2 | ||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">DNA( | + | <td width="257">DNA(digested sample)</td> |
<td>2</td> | <td>2</td> | ||
</tr> | </tr> | ||
Line 241: | Line 259: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>7</td> | <td>7</td> | ||
</tr> | </tr> | ||
Line 250: | Line 268: | ||
</table> | </table> | ||
- | + | -->37℃ 30in | |
- | + | ||
- | + | [[Team:Chiba/protocol/gelcheck|Gel Check]] | |
- | + | *[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2① | |
- | + | *[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2② | |
- | + | *[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](PCR Product)③ | |
+ | *[http://partsregistry.org/Part:BBa_E2050 BBa_E2050]④ | ||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">Sample No</td> | + | <td width="257">Sample No.</td> |
<td>①~②</td><td>③,④</td> | <td>①~②</td><td>③,④</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>DNA( | + | <td>DNA(digested sample)</td> |
<td>10</td><td>1</td> | <td>10</td><td>1</td> | ||
</tr> | </tr> | ||
Line 276: | Line 295: | ||
</table> | </table> | ||
- | + | *Luciferase check-->no light | |
+ | |||
+ | [[Team:Chiba/jk/γ/week 2|>Last Week]] | ||
- | + | [[Team:Chiba/jk/γ/week 4|>Next Week]] | |
- | + | ||
Latest revision as of 06:30, 24 October 2008
Contents |
WEEK 3
31,August,2008
- [http://partsregistry.org/Part:BBa_R0010 BBa_R0010]①
- PCR product([http://partsregistry.org/Part:BBa_J52008 BBa_J52008])②
Sample No. | ① | ② |
DNA | 10 | 10 |
SpeⅠ | 1 | 0 |
PstⅠ | 1 | 1 |
XbaⅠ | 0 | 1 |
BSA(x100) | 0.5 | 0.5 |
Buffer2 | 5 | 0 |
Buffer3 | 0 | 5 |
dH2O | 28.5 | 28.5 |
TOTAL | 50 | 50 |
-->37℃ 3hour
-->①20μl refine
-->②20μl refine
1,September,2008
- vector:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
- negative control:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]②
Sample No. | ① | ② |
vector | 2 | 2 |
insert | 3 | 0 |
Ligase Buffer | 2 | 2 |
Ligase | 1 | 1 |
dH2O | 3 | 5 |
TOTAL | 10 | 10 |
-->R/T 2hour
- [http://partsregistry.org/Part:BBa_R0079 BBa_R0079](Las promoter)
- [http://partsregistry.org/Part:BBa_R0071 BBa_R0071](RhlR promoter)
- [http://partsregistry.org/Part:BBa_R0077 BBa_R0077](cinR promoter+RBS)
- [http://partsregistry.org/Part:BBa_R0078 BBa_R0078](cinR promoter)
- [http://partsregistry.org/Part:BBa_R0062 BBa_R0062](Lux promoter)
2,September,2008
Colony PCR
- [http://partsregistry.org/Part:BBa_R0010 BBa_R0010]+[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
Sample No. | ① |
culture | 1 |
Fwd primer | 1.5 |
Rev primer | 1.5 |
Thermo pol Buffer | 3 |
dNTP mix | 3 |
Taq DNA pol (NEB) | 0.2 (1 unit) |
dH2O | 19.8 |
TOTAL | 30 |
- [http://partsregistry.org/Part:BBa_R0079 BBa_R0079]
- [http://partsregistry.org/Part:BBa_R0071 BBa_R0071]
- [http://partsregistry.org/Part:BBa_R0077 BBa_R0077]
- [http://partsregistry.org/Part:BBa_R0078 BBa_R0078]
- [http://partsregistry.org/Part:BBa_R0062 BBa_R0062]
3,September,2008
- [http://partsregistry.org/Part:BBa_R0079 BBa_R0079]①
- [http://partsregistry.org/Part:BBa_R0071 BBa_R0071]②
- [http://partsregistry.org/Part:BBa_R0077 BBa_R0077]③
- [http://partsregistry.org/Part:BBa_R0078 BBa_R0078]④
- [http://partsregistry.org/Part:BBa_R0062 BBa_R0062]⑤
Sample No. | ①~⑤ |
DNA tamplate | 5 |
Buffer3 | 1 |
dH2O | 4 |
EcoRⅠ | 0.1 |
TOTAL | 10 |
-->37℃30min
-->Gel check
Sample No. | ①~⑤ |
DNA(digestion sample) | 10 |
loading Dye | 2 |
TOTAL | 12 |
4,Septempber,2008
- Ligation product
- rLUciferase([http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] ) x3
- Venus YFP([http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]) x3
5,September,2008
- [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2
- [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2
DNA(digested sample) | 2 |
PstⅠ | 0.1 |
BSA | 0.8 |
Buffer3 | 1 |
dH2O | 7 |
TOTAL | 10 |
-->37℃ 30in
- [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2①
- [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]+[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] x2②
- [http://partsregistry.org/Part:BBa_J52008 BBa_J52008](PCR Product)③
- [http://partsregistry.org/Part:BBa_E2050 BBa_E2050]④
Sample No. | ①~② | ③,④ |
DNA(digested sample) | 10 | 1 |
loading Dye | 2 | 1 |
TOTAL | 12 | 6 |
- Luciferase check-->no light
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