Team:KIT-Kyoto/Notebook-week1p

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<td width="800px" height="510px" valign="top">
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<div id="MIGI">
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<h2>August 25th</h2>
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<br>
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<Table Border Cellspacing="0">
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<Tr><Td> All DNA </Td><Td> 500ng </Td><Td> 1ug </Td></Tr>
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<tr><td> pEGFP-C </td><td> 3uL </td><Td> 6uL </Td></tr>
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<Tr><Td> H buffer(TOYOBO) </Td><Td> 5uL </Td><Td> 5uL </Td></Tr>
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<tr><td> BamHⅠ(TOYOBO) </td><td> 1uL </td><Td> 1uL </Td></tr><tr>
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<td> BglⅡ(TOYOBO) </td><td> 1uL </td><Td> 1uL </Td></tr><tr>
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<td> 100×BSA </td><td> 0.5uL </td><Td> 0.5uL </Td></tr><tr>
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<td> dH<sub>2</sub>O </td><td> 39.5uL </td><Td> 36.5uL </Td></tr><tr>
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<td> Total </td><td> 50uL </td><Td> 50uL </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/d/d0/0825kit.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> DNA sample </td><td> 23uL </td></tr>
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<Tr><Td> 4 buffer(NEB) </Td><Td> 5uL </Td></Tr>
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<tr><td> EcoRⅠ-HF(NEB) </td><td> 1uL </td></tr><tr>
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<td> SpeⅠ(NEB) </td><td> 1.5uL </td></tr>
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<td> 100×BSA </td><td> 0.5uL </td></tr>
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<td> dH<sub>2</sub>O </td><td> 19uL </td></tr>
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<td> Total <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></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><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> 68.3uL </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>  58℃</td><td> 30min(UAS) or 2min10sec(pSB1C3)</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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<h2>August 26th</h2>
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<br>
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<img src="http://2012.igem.org/wiki/images/6/6d/0826.png" width="500" height="300">
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<br>
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<br>
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<h2>August 27th</h2>
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<br>
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<Table Border Cellspacing="0">
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<Tr><Td> pEGFP-C2(cut) </Td><Td> 1uL </Td></Tr>
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<tr><td> Ligation high </td><td> 2.5uL </td></tr><tr>
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<td> dH<sub>2</sub>O </td><td> 5uL </td></tr><tr>
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<td> Total <td> 7.5uL </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/e/e6/0826bkit.png" width="500" height="300">
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<br><br>
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<img src="http://2012.igem.org/wiki/images/9/99/0826ckit.png" width="500" height="300">
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<br><br>
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<h2>August 28th</h2>
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<br>
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<h2>August 29th</h2>
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<br>
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<Table Border Cellspacing="0">
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<Tr><Td> pEGFP-C2-1, -2, -3, -4 </Td><Td> 1uL </Td></Tr>
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<tr><td> 10×H buffer(TOYOBO) </td><td> 0.5uL </td></tr><tr>
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<tr><td> XhoⅠ </td><td> 0.1uL </td></tr><tr>
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<tr><td> PstⅠ </td><td> 0.1uL </td></tr><tr>
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<td> dH<sub>2</sub>O </td><td> 3.3uL </td></tr><tr>
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<td> Total <td> 5uL </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/9/97/0828kit.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> 0.5uL </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>
 +
<td> 68℃ </td><td> 30sec(UAS) or 1min(EGFP) </td><Td> 25 cycle </Td></tr>
 +
<tr><td> 68℃ </td><td> 30sec(UAS) or 1min(EGFP) </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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<img src="http://2012.igem.org/wiki/images/d/d2/0829b.png" width="500" height="300">
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<br><br>
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Revision as of 09:57, 25 September 2012






August 25th


 All DNA  500ng  1ug 
 pEGFP-C  3uL  6uL 
 H buffer(TOYOBO)  5uL  5uL 
 BamHⅠ(TOYOBO)  1uL  1uL 
 BglⅡ(TOYOBO)  1uL  1uL 
 100×BSA  0.5uL  0.5uL 
 dH2O  39.5uL  36.5uL 
 Total  50uL  50uL 




 DNA sample  23uL 
 4 buffer(NEB)  5uL 
 EcoRⅠ-HF(NEB)  1uL 
 SpeⅠ(NEB)  1.5uL 
 100×BSA  0.5uL 
 dH2O  19uL 
 Total  50uL 


 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  Circle 
  95℃ 2min
  95℃ 15sec 25 cycle 
  58℃ 30min(UAS) or 2min10sec(pSB1C3) 25 cycle 
  68℃ 2min30sec 25 cycle 
  68℃ 2min30sec
  14℃ ∞


August 26th




August 27th


 pEGFP-C2(cut)  1uL 
 Ligation high  2.5uL 
 dH2O  5uL 
 Total  7.5uL 






August 28th


August 29th


 pEGFP-C2-1, -2, -3, -4  1uL 
 10×H buffer(TOYOBO)  0.5uL 
 XhoⅠ  0.1uL 
 PstⅠ  0.1uL 
 dH2O  3.3uL 
 Total  5uL 




 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(UAS) or 1min(EGFP)  25 cycle 
 68℃  30sec(UAS) or 1min(EGFP) 
 14℃  ∞