Team:ZJU-China/Notebook/September

From 2011.igem.org

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<p>&nbsp;</p>
<p>&nbsp;</p>
</div>
</div>
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<div class="block" id="nsheet">
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<h3>Week11</h3><hr/>
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<table id="notesheet" width="650" border="1" cellspacing="0" cellpadding="1">
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  <tr><td width="76">Day</td><td width="349">Note</td></tr>
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  <tr>
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    <td id="sheetleft">Sept.12th
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Monday
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</td>
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    <td><table id="intable" width="603" border="0" cellspacing="0" cellpadding="1">
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    <td width="191">• Colony PCR: 1A3, 1K3, lig RFP+YFP+1C3.
 +
 +
</td>
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 +
 
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    <td  >• Run the PCR results.
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 +
</td>
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<td>• Miniprep: lig RFP+YFP+1C3
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</td>
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 +
</table></td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Sept.13th
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Tuesday
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 +
 +
 +
 +
 +
 +
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</td>
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    <td><table id="intable" width="615" border="0" cellspacing="0" cellpadding="1">
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  <td>• Transform 2M, 2I, 6I, 23L.</td>
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  <td>• Purification: 1K3</td>
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 +
</table></td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Sept.14th
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Wednesday
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 +
 +
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</td>
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    <td><table width="640" border="0" cellspacing="0" cellpadding="1">
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  <tr>
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  <td>• Miniprep: 20H, 20J, 22B</td>
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  <td>• Cut: 1K3 with E+P, lig3 with S+E, 12I(CFP) with P+X,</td>
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  <td>• Gel excision and /or purification: 1K3, lig3, 12I(CFP)</td>
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  <td>• Run the gel.<br/>
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• Culture: 1I, 1K, 3L, 5E, 7C
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</td>
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<td>• Miniprep: RFP+YFP+1C3.<br/>
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• Run the gel of 1K3, lig3, CFP again.
 +
</td>
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<td>• Gel excision and purification<br/>
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• Ligation: Lig3+CFP+1K3
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</td>
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  </tr>
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</table>
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</td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Sept.15th
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Thursday
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 +
 +
 +
</td>
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    <td><table width="618" border="0" cellspacing="0" cellpadding="1">
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  <tr>
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<td>• Preserve: 1I, 1K, 3C, 5E, 7C<br/>
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• Transform the ligation results
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</td>
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<td>• Colony PCR: 2M, 2I，6I，23L<br/>
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• Cut: 20H, 20J, 22B with X+P
 +
 +
</td>
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<td>• Miniprep: 2M-1, 2I-1, 6I-1, 23L-1<br/>
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• Cut: 2M, 2I with E+S
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 +
 +
</td>
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  </tr>
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</table>
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</td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Sept.16th
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Friday
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 +
 +
 +
 +
</td>
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    <td><table width="627" border="0" cellspacing="0" cellpadding="1">
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      <tr>
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      <td >• Colony PCR: RFP+YFP+1K3</td>
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      <td>• Cut 13K with E+S, 10I with P+X, 1C3 with E+P
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</td>
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<td>• Run the gel: 13K, 10I, 1C3, 2I, 2M, 20J, 22H, 22B</td>
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<td>• Ligation: 13K+10I+1C3<br/>
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• Culture: YFP(Amp), CFP(Amp)
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</td>
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      </tr>
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    </table></td>
 +
  </tr>
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  <tr>
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    <td id="sheetleft">Sept.17th
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Saturday
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 +
 +
 +
 +
 +
</td>
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    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
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  <tr>
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    <td>• Order primers for 20H, 20J<br/>
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• Culture 20H-1, 20J-1, 22B-1
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</td>
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  <td>• Plate the ligation results<br/>
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• Culture RFP+YFP+CFP+1K3
 +
 +
</td>
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<td>• Colony PCR: 3A, 1C3<br/>
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• Preserve: 1K3, RFP+YFP+1C3.
 +
 +
</td>
 +
  </tr>
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</table>
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</td>
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  </tr>
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  <tr>
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    <td id="sheetleft">Sept.18th
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Sunday
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 +
 +
</td>
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    <td><table width="620" border="0" cellspacing="0" cellpadding="1">
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      <tr>
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        <td width="244">• Miniprep: 20J, 20H, RFP+YFP+CFP+1K3
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1A3
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</td>
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<td width="164">• Run the gel of PCR results
 +
 +
</td>
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<td>• Cut RFP+YFP+CFP+1K3
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with E+P<br/>
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• Colony PCR: 13K+10I+1C3
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</td>
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<td>• PCR: fdhF+rush+Vgb+tetR<br/>
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• Preserve: 20H, 20J, 22B
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</td>
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<td>• Run the gel: 20H, 20J, 22B<br/>
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• Cut the bands and purification.
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</td>
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<td>• Run the gel: 13K+10I+1C3, RFP+YFP+CFP+1K3<br/>
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• Culture: 13K+10I+1C3<br/>
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• Grad PCR: fdhF.
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</td>
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      </tr>
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    </table></td>
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  </tr>
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</table>
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<p>&nbsp;</p>
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</div>
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Revision as of 06:16, 3 October 2011

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Labnote

 Biobrick group | Biofilm group 

This group...........

Week9

DayNote
Aug.29th Monday
• Meeting. Summarize all the experiment work so far. • Cut: 22M(X+P), 1C3(E+P)
Aug.30th Tuesday
Aug.31st Wednesday
• Cut vgb+YFP+tetR, fdhF+RFP+tetR.
• Run the digestion results		 • Culture the positive results
Sept.1st Thursday
• Miniprep: Y#4, R#2/3/4/5
• Cut Y#4, R#2/3/4/5 		•Run the gel.
• Send R#3,Y#4 sample to sequencing 		• Hypoxia culture: RFP3, RFP4
• Preserve Y#4, R#2/3/4/5
Sept.2nd Friday
• Phusion PCR • Hypoxia Culture: R, Y
• Run the PCR results. Bands are confirmed right. 		• Purification: vgb, YFP, tetR.
Sept.3rd Saturday
• Culture: YFP, RFP, A25
• Culture in micro-oxygen: YFP, RFP, A25 		• Culture in slope medium: RFP, YFP, A25
• Hypoxia culture: RFP, YFP, A25 		• Check the fluorescence.
• Neither RFP in hypoxia or oxygen condition. Little YFP both in hypoxia and oxygen.
Sept.4th Sunday
• Miniprep: RFP3, YFP4 • Cut: RFP3, YFP4, !C3.

 

Week10

DayNote
Sept.5th Monday
• Run the gel: Yu’s PCR results, 1C3, RFP3, YFP4
• Purification: RFP3, YFP4 		• Ligation: RFP3+1C3, YFP4+1C3
• Hypoxia culture: RFP3, YFP4, A25. 		• Transform the ligation results.
• Confirmed RFP expression only in hypoxia condition.
Sept.6th Tuesday
• Check the RFP, YFP plates. • Colony PCR
Sept.7th Wednesday
• Culture YFP1/2, RFP3/4
Sept.8th Thurday
• Preserve the strain of RFP1, YFP1/2 in 1C3.
• Miniprep the remains. 		• Cut: YFP1/2, RFP1 • Run the digestion results.
• Culture RFP2
Sept.9th Friday
• Miniprep: RFP2, YFP2, RFP1 • Cut them and run the gel.
Sept.10th Saturday
• Cut YFP(1A3), RFP(1A3) with S+E • Purification: RFP(1A3), YFP(1A3) • Ligation: RFP+1C3, YFP+1C3 • Transform the ligation results.
Sept.11th Sunday
• Check the plates. • Culture lig RFY+YFP+IC3
• Culture 1K3 		• Colony PCR: RFP(1C3)， YFP(1C3)

 

Week11

DayNote
Sept.12th Monday
• Colony PCR: 1A3, 1K3, lig RFP+YFP+1C3. • Run the PCR results. • Miniprep: lig RFP+YFP+1C3
Sept.13th Tuesday
• Transform 2M, 2I, 6I, 23L. • Purification: 1K3
Sept.14th Wednesday
• Miniprep: 20H, 20J, 22B • Cut: 1K3 with E+P, lig3 with S+E, 12I(CFP) with P+X, • Gel excision and /or purification: 1K3, lig3, 12I(CFP) • Run the gel.
• Culture: 1I, 1K, 3L, 5E, 7C 		• Miniprep: RFP+YFP+1C3.
• Run the gel of 1K3, lig3, CFP again. 		• Gel excision and purification
• Ligation: Lig3+CFP+1K3
Sept.15th Thursday
• Preserve: 1I, 1K, 3C, 5E, 7C
• Transform the ligation results 		• Colony PCR: 2M, 2I，6I，23L
• Cut: 20H, 20J, 22B with X+P 		• Miniprep: 2M-1, 2I-1, 6I-1, 23L-1
• Cut: 2M, 2I with E+S
Sept.16th Friday
• Colony PCR: RFP+YFP+1K3 • Cut 13K with E+S, 10I with P+X, 1C3 with E+P • Run the gel: 13K, 10I, 1C3, 2I, 2M, 20J, 22H, 22B • Ligation: 13K+10I+1C3
• Culture: YFP(Amp), CFP(Amp)
Sept.17th Saturday
• Order primers for 20H, 20J
• Culture 20H-1, 20J-1, 22B-1 		• Plate the ligation results
• Culture RFP+YFP+CFP+1K3 		• Colony PCR: 3A, 1C3
• Preserve: 1K3, RFP+YFP+1C3.
Sept.18th Sunday
• Miniprep: 20J, 20H, RFP+YFP+CFP+1K3 1A3 • Run the gel of PCR results • Cut RFP+YFP+CFP+1K3 with E+P
• Colony PCR: 13K+10I+1C3 		• PCR: fdhF+rush+Vgb+tetR
• Preserve: 20H, 20J, 22B 		• Run the gel: 20H, 20J, 22B
• Cut the bands and purification. 		• Run the gel: 13K+10I+1C3, RFP+YFP+CFP+1K3
• Culture: 13K+10I+1C3
• Grad PCR: fdhF.

 

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