Team:Chiba/jk/γ/week 2
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 2 == | == WEEK 2 == | ||
== 24,August 2008 == | == 24,August 2008 == | ||
+ | [[Team:Chiba/protocol/transformation|Transformation]] | ||
+ | *[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](yeast optimized YFP)-->colony | ||
+ | |||
+ | [[Team:Chiba/protocol/digestion|Digestion]] | ||
+ | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)① | ||
+ | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)② | ||
+ | *Menbren Venus YFP③ | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
<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>③</td> | <td>①</td><td>②</td><td>③</td> | ||
</tr> | </tr> | ||
Line 26: | Line 28: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>6</td><td>6</td><td>6</td> | <td>6</td><td>6</td><td>6</td> | ||
</tr> | </tr> | ||
Line 38: | Line 40: | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
+ | |||
+ | -->37℃ 30min | ||
+ | |||
+ | |||
<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(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td> | <td>①(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td> | ||
</tr> | </tr> | ||
Line 57: | Line 63: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | |||
+ | |||
<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>③</td> | <td>①</td><td>②</td><td>③</td> | ||
</tr> | </tr> | ||
Line 71: | Line 79: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>3</td><td>3</td><td>3</td> | <td>3</td><td>3</td><td>3</td> | ||
</tr> | </tr> | ||
Line 79: | Line 87: | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
+ | -->135V 30min | ||
== 25,August,2008 == | == 25,August,2008 == | ||
- | + | [[Team:Chiba/protocol/PCR|PCR]] | |
- | + | *[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]① | |
- | + | *[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]② | |
- | + | *BL21③ | |
- | + | *[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④ | |
- | + | *[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]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</td><td>②</td><td>③</td><td>④</td><td>⑤</td> | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td> | ||
</tr> | </tr> | ||
Line 120: | Line 130: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>68</td><td>68</td><td>68</td><td>68</td><td>68</td> | <td>68</td><td>68</td><td>68</td><td>68</td><td>68</td> | ||
</tr> | </tr> | ||
Line 128: | Line 138: | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
+ | -->[[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</td><td>②</td><td>③</td><td>④</td><td>⑤</td> | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td> | ||
</tr> | </tr> | ||
Line 143: | Line 156: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td> | <td>4</td><td>4</td><td>4</td><td>4</td><td>4</td> | ||
</tr> | </tr> | ||
Line 152: | Line 165: | ||
</table> | </table> | ||
- | + | [[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]] | |
- | + | *Venus | |
- | + | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | |
- | + | ||
- | + | ||
- | + | [[Team:Chiba/protocol/transformation|Transformation]] | |
- | + | *Venus | |
- | + | *PUC19 | |
- | + | *pGFPUV | |
- | + | *pGEX Venus YFP | |
- | + | *Membren Venus YFP | |
+ | [[Team:Chiba/protocol/PCR|PCR]] | ||
+ | *[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]① | ||
<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 193: | Line 208: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>68</td> | <td>68</td> | ||
</tr> | </tr> | ||
Line 204: | Line 219: | ||
== 26,August,2008 == | == 26,August,2008 == | ||
- | + | [[Team:Chiba/protocol/gelcheck|Gel Check]] | |
- | + | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | |
- | + | *[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | |
- | + | *[http://partsregistry.org/Part:BBa_J63001 BBa_J63001] | |
<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>③</td><td>④</td><td>⑤</td> | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td> | ||
</tr> | </tr> | ||
Line 222: | Line 237: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td> | <td>4</td><td>4</td><td>4</td><td>4</td><td>4</td> | ||
</tr> | </tr> | ||
Line 231: | Line 246: | ||
</table> | </table> | ||
- | + | [[Team:Chiba/protocol/digestion|Digestion]] | |
- | + | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | |
+ | |||
<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>BBa_F2620</td> | + | <td>[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 263: | Line 279: | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
+ | *insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008],[http://partsregistry.org/Part:BBa_E2030 BBa_E2030] | ||
+ | |||
<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> | + | <td>[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]</td><td>[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 298: | Line 316: | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | <br>->37℃ 3hour | + | <br>-->37℃ 3hour |
- | <br>->SAP | + | <br>-->SAP |
- | <br>->37℃ 30min | + | <br>-->37℃ 30min |
- | <br>->65℃ 15min | + | <br>-->65℃ 15min |
- | <br>->Zymo Clean | + | <br>-->[[Team:Chiba/protocol/DNA_Purification/zymocleam|Zymo Clean]] |
- | + | ||
- | + | [[Team:Chiba/protocol/ligation/ligation|Ligation]] | |
- | + | ||
- | + | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]① | |
+ | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]② | ||
+ | *Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③ | ||
+ | |||
<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>③</td> | <td>①</td><td>②</td><td>③</td> | ||
</tr> | </tr> | ||
Line 329: | Line 350: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>0</td><td>2.5</td><td>10</td> | <td>0</td><td>2.5</td><td>10</td> | ||
</tr> | </tr> | ||
Line 337: | Line 358: | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ||
- | + | -->RT 2hour | |
- | + | ||
- | + | -->[[Team:Chiba/protocol/transformation|Transformation]](XL10G) | |
- | + | ||
+ | -->37℃ over night | ||
+ | |||
+ | -->colony PCR | ||
+ | |||
+ | -->[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]] | ||
== 28,August,2008 == | == 28,August,2008 == | ||
- | + | ||
+ | [[Team:Chiba/jk/γ/Trl|Time Responce(liquid)]] | ||
Line 351: | Line 378: | ||
== 29,August,2008 == | == 29,August,2008 == | ||
- | + | ||
+ | [[Team:Chiba/jk/γ/trs|TIME RESPONCE(solid)]] | ||
== 30,August,2008 == | == 30,August,2008 == | ||
- | + | [[Team:Chiba/protocol/transformation|Transformation]] | |
- | + | ||
- | + | *mcherry | |
- | + | ||
+ | [[Team:Chiba/protocol/PCR|PCR]] | ||
+ | |||
+ | *[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](rLuc) | ||
+ | |||
<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>BBa_J52008</td> | + | <td>[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 390: | Line 422: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>dH<sub>2</sub>O</td> |
<td>34</td> | <td>34</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
<td>50</td> | <td>50</td> | ||
Line 402: | Line 435: | ||
[[Team:Chiba/jk/γ/week 1|>Last week]] | [[Team:Chiba/jk/γ/week 1|>Last week]] | ||
- | + | ||
+ | [[Team:Chiba/jk/γ/week 3|>Next week]] | ||
Latest revision as of 05:50, 24 October 2008
Contents |
WEEK 2
24,August 2008
- [http://partsregistry.org/Part:BBa_J63001 BBa_J63001](yeast optimized YFP)-->colony
- [http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)①
- [http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)②
- Menbren Venus YFP③
Sample No. | ① | ② | ③ |
DNA tamplate | 3 | 3 | 3 |
Buffer3 | 1 | 1 | 1 |
dH2O | 6 | 6 | 6 |
EcoRⅠ | 0.5 | 0.5 | 0.5 |
TOTAL | 10 | 10 | 10 |
-->37℃ 30min
Sample No. | ①(afetr digestion) | ②(after digestion) | ③(afetr digestion) |
DNA tamplate | 10 | 10 | 10 |
loading Dye | 2 | 2 | 2 |
TOTAL | 12 | 12 | 12 |
Sample No. | ① | ② | ③ |
DNA tamplate | 2 | 2 | 2 |
loading Dye | 1 | 1 | 1 |
dH2O | 3 | 3 | 3 |
TOTAL | 6 | 6 | 6 |
-->135V 30min
25,August,2008
- [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
- [http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
- BL21③
- [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④
- [http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤
Sample No. | ① | ② | ③ | ④ | ⑤ |
DNA tamplate | 1 | 1 | 1 | 1 | >1 |
FW primer | 5(Venus_YFP_fwd) | 5(LacZ_fwd) | 5(LacZ_fwd) | 5(rLUC_fwd) | 5(fLuc_fwd) |
VR primer | 5(VR) | 5(LacZ_rev) | 5(LacZ_rev) | 5(VR) | 5(VR) |
dNTPmix | 10 | 10 | 10 | 10 | 10 |
Thermo pol buffer | 10 | 10 | 10 | 10 | 10 |
DNA pol(VENT) | 1 | 1 | 1 | 1 | 1 |
dH2O | 68 | 68 | 68 | 68 | 68 |
TOTAL | 100 | 100 | 100 | 100 | 100 |
-->Gel Check
Sample No. | ① | ② | ③ | ④ | ⑤ |
DNA tamplate | 1 | 1 | 1 | 1 | >1 |
loading Dye | 1 | 1 | 1 | 1 | 1 |
dH2O | 4 | 4 | 4 | 4 | 4 |
TOTAL | 6 | 6 | 6 | 6 | 6 |
- Venus
- [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
- Venus
- PUC19
- pGFPUV
- pGEX Venus YFP
- Membren Venus YFP
- [http://partsregistry.org/Part:BBa_J63001 BBa_J63001]①
Sample No. | ① |
DNA tamplate | 1 |
FW primer(Venus) | 5 |
VR primer | 5 |
dNTPmix | 10 |
Thermo pol buffer | 10 |
DNA pol(VENT) | 1 |
dH2O | 68 |
TOTAL | 100 |
26,August,2008
- [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
- [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
- [http://partsregistry.org/Part:BBa_J63001 BBa_J63001]
Sample No. | ① | ② | ③ | ④ | ⑤ |
DNA tamplate | 1 | 1 | 1 | 1 | >1 |
Loading Dye | 1 | 1 | 1 | 1 | 1 |
dH2O | 4 | 4 | 4 | 4 | 4 |
TOTAL | 6 | 6 | 6 | 6 | 6 |
- vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
Sample No. | [http://partsregistry.org/Part:BBa_F2620 BBa_F2620] |
DNA tamplate | 100 |
SpeⅠ | 2 |
PstⅠ | 2 |
BSA(x10) | 13 |
Buffer2 | 13 |
TOTAL | 130 |
- insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008],[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
Sample No. | [http://partsregistry.org/Part:BBa_J52008 BBa_J52008] | [http://partsregistry.org/Part:BBa_E2030 BBa_E2030] |
DNA tamplate | 30 | 30 |
XbaⅠ | 1 | 1 |
PstⅠ | 1 | 1 |
DpnⅠ | 1 | 1 |
Buffer3 | 4 | 4 |
BSA(x10) | 4 | 4 |
TOTAL | 40 | 40 |
-->37℃ 3hour
-->SAP
-->37℃ 30min
-->65℃ 15min
-->Zymo Clean
- vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
- vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]②
- Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③
Sample No. | ① | ② | ③ |
vector | 5 | 5 | 5 |
inser | 10 | 7.5 | 0 |
Ligase Buffer | 4 | 4 | 4 |
Ligase | 1 | 1 | 1 |
dH2O | 0 | 2.5 | 10 |
TOTAL | 20 | 20 | 20 |
-->RT 2hour
-->Transformation(XL10G)
-->37℃ over night
-->colony PCR
-->Mini prep
28,August,2008
29,August,2008
30,August,2008
- mcherry
- [http://partsregistry.org/Part:BBa_J52008 BBa_J52008](rLuc)
Sample No. | [http://partsregistry.org/Part:BBa_J52008 BBa_J52008] |
DNA tamplate | 1 |
rLuc_fwd | 2.5 |
Rev primer | 2.5 |
Thermo pol Buffer | 5 |
dNTPmix | 5 |
Vent pol | 0.5 |
dH2O | 34 |
TOTAL | 50 |
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