Team:KIT-Kyoto/Notebook-week2p

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<td width="800px" height="510px" valign="top">
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<div id="MIGI">
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<h2>August 31st</h2>
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<br>
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<Table Border Cellspacing="0">
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<Tr><Td></Td><Td> All samples </Td></Tr>
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<Tr><Td> DNA template </Td><Td> 0.5uL </Td></Tr>
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<tr><td> 10× KOD plus buffer </td><td> 10uL </td></tr>
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<Tr><Td> 2mM dNTPs </Td><Td> 10uL </Td></Tr>
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<Tr><Td> 25mM MgSO4 </Td><Td> 3.2uL </Td></Tr>
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<td> 10P 5’ primer </td><td> 3uL </td></tr>
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<Tr><td> 10P 3’ primer </td><td> 3uL </td></tr>
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<Tr><td> KOD plus </td><td> 2uL </td></tr>
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<Tr><td> dH<sub>2</sub>O </td><td> 68.3uL </td></tr>
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<Tr><td> Total </td><td> 100uL </td></tr>
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</Table>
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<br><BR>
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<Table Border Cellspacing="0">
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<Tr><Td> Temperature</Td><Td> Time </Td><Td>Cycle</Td></Tr>
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<tr><td>  95℃ </td><td> 2min </td><Td></Td></tr>
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<Tr><Td>  95℃ </Td><Td> 15sec </Td><Td> 25 cycle </Td></Tr>
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<tr><td>  58℃ </td><td> 30sec </td><Td> 25 cycle </Td></tr><tr>
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<td>  68℃ </td><td> 30sec </td><Td> 25 cycle </Td></tr>
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<tr><td>  68℃ </td><td> 30sec </td><Td></Td></tr>
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<tr><td>  14℃ </td><td> ∞ </td><Td></Td></tr>
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</Table>
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<br><BR>
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<h2>September 1st</h2>
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<br>
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<img src="http://2012.igem.org/wiki/images/b/b0/0901akit.png" width="500" height="300">
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<br><br>
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<Table Border Cellspacing="0">
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<Tr><Td></Td><Td> All samples </Td></Tr>
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<tr><td> DNA template </td><td> 0.3uL </td></tr>
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<Tr><Td> 10× KOD plus buffer </Td><Td> 10uL </Td></Tr>
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<tr><td> 2mM dNTPs </td><td> 10uL </td></tr><tr>
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<td> 25mM MgSO4 </td><td> 3.2uL </td></tr>
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<td> 10P 5’ primer </td><td> 3uL </td></tr>
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<td> 10P 3’ primer </td><td> 3uL </td></tr>
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<td> KOD plus </td><td> 2uL </td></tr>
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<td> dH<sub>2</sub>O </td><td> 68.5uL </td></tr>
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<td> Total <td> 100uL </td></tr>
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</Table>
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<br><BR>
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<Table Border Cellspacing="0">
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<Tr><Td> Temperature </Td><Td> Time </Td><Td> Circle </Td></Tr>
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<tr><td>  95℃ </td><td> 2min </td><Td></Td></tr>
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<Tr><Td>  95℃ </Td><Td> 15sec </Td><Td> 25 cycle </Td></Tr>
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<tr><td>  59℃ </td><td> 30sec </td><Td> 25 cycle </Td></tr><tr>
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<td>  68℃ </td><td> 1min30sec(Act5c and LacZ) or 2min(GAL4 and U-T) </td><Td> 25 cycle </Td></tr><tr>
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<td>  68℃ </td><td> 1min30sec(Act5c and LacZ) or 2min(GAL4 and U-T) </td><Td></Td></tr><tr>
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<td>  14℃ </td><td> ∞ </td><Td></Td></tr>
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</Table>
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<br><br>
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<img src="http://2012.igem.org/wiki/images/6/67/0901bkit.png" width="500" height="300">
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<br><br>
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<h2>September 3rd</h2>
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<BR>
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<Table Border Cellspacing="0">
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<Tr><Td></Td><Td> All samples </Td></Tr>
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<tr><td> DNA template </td><td> 0.2uL </td></tr>
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<Tr><Td> 10× KOD plus buffer </Td><Td> 5uL </Td></Tr>
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<tr><td> 2mM dNTPs </td><td> 5uL </td></tr><tr>
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<td> 25mM MgSO4 </td><td> 1.6uL </td></tr><tr>
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<td> 10P 5’ primer </td><td> 1.5uL </td></tr><tr>
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<td> 10P 3’ primer </td><td> 1.5uL </td></tr><tr>
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<td> KOD plus </td><td> 1u L</td></tr><tr>
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<td> dH<sub>2</sub>O </td><td> 34.2uL </td></tr><tr>
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<td> Total <td> 50uL </td></tr>
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</Table>
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<br><BR>
 +
<Table Border Cellspacing="0">
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<Tr><Td> Temperature </Td><Td> Time </Td><Td> Circle </Td></Tr>
 +
<tr><td> 95℃ </td><td> 2min </td><Td></Td></tr>
 +
<Tr><Td> 95℃ </Td><Td> 15sec </Td><Td> 25 cycle </Td></Tr>
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<tr><td> 55℃(GAL4) and 58℃(Except for GAL4) </td><td> 30sec </td><Td> 25 cycle </Td></tr><tr>
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<td> 68℃ </td><td> 2min30sec </td><Td> 25 cycle </Td></tr>
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<td> 68℃ </td><td> 2min30sec </td><Td></Td></tr>
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<td> 14℃ </td><td> ∞ </td><Td></Td></tr>
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</Table>
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<br><br>
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<img src="http://2012.igem.org/wiki/images/c/ca/0903kit.png" width="500" height="300">
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<br><br>
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<h2>September 4th</h2>
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<br>
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<Table Border Cellspacing="0">
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<Tr><Td> 9/3 GAL4 </Td><Td> 40uL </Td></Tr>
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<tr><td> M buffer(TOYOBO) </td><td> 5uL </td></tr>
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<Tr><Td> XbaⅠ(TOYOBO) </Td><Td> 0.5uL </Td></Tr>
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<tr><td> SpeⅠ(TOYOBO) </td><td> 0.5uL </td></tr><tr>
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<td> dH<sub>2</sub>O </td><td> 4uL </td></tr><tr>
 +
<td> Total </td><td> 50uL </td></tr>
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</Table>
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<br><BR>
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<Table Border Cellspacing="0">
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<Tr><Td> each DNA </Td><Td> 40uL </Td></Tr>
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<tr><td> 3 buffer(NEB) </td><td> 5uL </td></tr>
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<Tr><Td> BglⅡ(NEB) </Td><Td> 0.5uL </Td></Tr><tr>
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<td> dH<sub>2</sub>O </td><td> 4.5uL </td></tr><tr>
 +
<td> Total </td><td> 50uL </td></tr>
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</Table>
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<br><BR>
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 +
<Table Border Cellspacing="0">
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<Tr><Td> 9/3 GAL4 </Td><Td> 23uL </Td></Tr>
 +
<tr><td> M buffer(TOYOBO) </td><td> 10uL </td></tr>
 +
<Tr><Td> XbaⅠ(TOYOBO) </Td><Td> 1uL </Td></Tr>
 +
<Tr><Td> SpeⅠ(TOYOBO) </Td><Td> 1uL </Td></Tr>
 +
<Tr><Td> CIP </Td><Td> 0.5uL </Td></Tr>
 +
<td> dH<sub>2</sub>O </td><td> 64.5uL </td></tr>
 +
<td> Total </td><td> 100uL </td></tr>
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</Table>
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<br><BR>
 +
 +
<img src="http://2012.igem.org/wiki/images/b/b0/0904kit.png" width="500" height="300">
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<br><br>
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<h2>September 5th</h2>
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<br>
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<Table Border Cellspacing="0">
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<Tr><Td></Td><td> EGFP </td><Td> pSB1C3 </Td></Tr>
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<tr><td> DNA template </td><td> 40uL </td><Td> 23uL </Td></tr>
 +
<Tr><Td> 3 buffer </Td><Td> 5uL </Td><Td> 5uL </Td></Tr>
 +
<tr><td> BglⅡ </td><td> 0.5uL </td><Td> 0.5uL </Td></tr><tr>
 +
<td> dH<sub>2</sub>O </td><td> 4.5uL </td><Td> 21.5uL </Td></tr>
 +
<td> Total </td><td> 50uL </td><Td> 50uL </Td></tr>
 +
</Table>
 +
<br><BR>
 +
 +
<img src="http://2012.igem.org/wiki/images/3/3a/0905akit.png" width="500" height="300">
 +
<br><br>
 +
<Table Border Cellspacing="0">
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<Tr><Td> DNA template </Td><Td> 40uL </Td></Tr>
 +
<tr><td> 4 buffer(NEB) </td><td> 5uL </td></tr>
 +
<Tr><Td> SpeⅠ </Td><Td> 0.5uL </Td></Tr>
 +
<Tr><Td> 100×BSA </Td><Td> 0.5uL </Td></Tr>
 +
<Tr><Td> CIP </Td><Td> 0.5uL </Td></Tr>
 +
<td> dH<sub>2</sub>O </td><td> 4uL </td></tr>
 +
<td> Total </td><td> 50uL </td></tr>
 +
</Table>
 +
<br><BR>
 +
 +
<Table Border Cellspacing="0">
 +
<Tr><Td></Td><Td> 1uL </Td></Tr>
 +
<tr><td></td><td> 1uL </td></tr>
 +
<Tr><Td> dH<sub>2</sub>O </Td><Td> 3uL </Td></Tr>
 +
<Tr><Td> Ligation high </Td><Td> 2.5uL </Td></Tr>
 +
<td> Total </td><td> 7.5uL </td></tr>
 +
</Table>
 +
<br><BR>
 +
<img src="http://2012.igem.org/wiki/images/6/63/0905bkit.png" width="500" height="300">
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<br><br>
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</div>
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</table>
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</td></tr>
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Revision as of 10:02, 25 September 2012






August 31st


 All samples 
 DNA template  0.5uL 
 10× KOD plus buffer  10uL 
 2mM dNTPs  10uL 
 25mM MgSO4  3.2uL 
 10P 5’ primer  3uL 
 10P 3’ primer  3uL 
 KOD plus  2uL 
 dH2O  68.3uL 
 Total  100uL 


 Temperature Time Cycle
  95℃  2min 
  95℃  15sec  25 cycle 
  58℃  30sec  25 cycle 
  68℃  30sec  25 cycle 
  68℃  30sec 
  14℃  ∞ 


September 1st




 All samples 
 DNA template  0.3uL 
 10× KOD plus buffer  10uL 
 2mM dNTPs  10uL 
 25mM MgSO4  3.2uL 
 10P 5’ primer  3uL 
 10P 3’ primer  3uL 
 KOD plus  2uL 
 dH2O  68.5uL 
 Total  100uL 


 Temperature  Time  Circle 
  95℃  2min 
  95℃  15sec  25 cycle 
  59℃  30sec  25 cycle 
  68℃  1min30sec(Act5c and LacZ) or 2min(GAL4 and U-T)  25 cycle 
  68℃  1min30sec(Act5c and LacZ) or 2min(GAL4 and U-T) 
  14℃  ∞ 




September 3rd


 All samples 
 DNA template  0.2uL 
 10× KOD plus buffer  5uL 
 2mM dNTPs  5uL 
 25mM MgSO4  1.6uL 
 10P 5’ primer  1.5uL 
 10P 3’ primer  1.5uL 
 KOD plus  1u L
 dH2O  34.2uL 
 Total  50uL 


 Temperature  Time  Circle 
 95℃  2min 
 95℃  15sec  25 cycle 
 55℃(GAL4) and 58℃(Except for GAL4)  30sec  25 cycle 
 68℃  2min30sec  25 cycle 
 68℃  2min30sec 
 14℃  ∞ 




September 4th


 9/3 GAL4  40uL 
 M buffer(TOYOBO)  5uL 
 XbaⅠ(TOYOBO)  0.5uL 
 SpeⅠ(TOYOBO)  0.5uL 
 dH2O  4uL 
 Total  50uL 


 each DNA  40uL 
 3 buffer(NEB)  5uL 
 BglⅡ(NEB)  0.5uL 
 dH2O  4.5uL 
 Total  50uL 


 9/3 GAL4  23uL 
 M buffer(TOYOBO)  10uL 
 XbaⅠ(TOYOBO)  1uL 
 SpeⅠ(TOYOBO)  1uL 
 CIP  0.5uL 
 dH2O  64.5uL 
 Total  100uL 




September 5th


 EGFP  pSB1C3 
 DNA template  40uL  23uL 
 3 buffer  5uL  5uL 
 BglⅡ  0.5uL  0.5uL 
 dH2O  4.5uL  21.5uL 
 Total  50uL  50uL 




 DNA template  40uL 
 4 buffer(NEB)  5uL 
 SpeⅠ  0.5uL 
 100×BSA  0.5uL 
 CIP  0.5uL 
 dH2O  4uL 
 Total  50uL 


 1uL 
 1uL 
 dH2O  3uL 
 Ligation high  2.5uL 
 Total  7.5uL