Team:IPN-UNAM-Mexico/Notebook/August 10 2009

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                                  <td><p>&nbsp;</p>
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                                    <p><strong>August 10th, 2009</strong></p>
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                                    <p>Calculus for Mix; plasmid linearization.</p>
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                                    <p>For 20 reactions.</p>
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                                    <p align="left" class="style8">&nbsp;</p>
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                                    <table width="24%" border="0" align="left" cellpadding="0" cellspacing="0">
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                                      <tr>
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                                        <td>&nbsp;</td>
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                                      </tr>
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                                    </table>
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                                    <table width="54%" border="1" align="left" cellpadding="0" cellspacing="0">
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                                      <tr>
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                                        <td class="style8"><div align="center" class="style11">Mix</div></td>
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                                        <td class="style11"><div align="center" class="style11">For every Eppendorf tube</div></td>
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                                        <td class="style11"><div align="center" class="style11">For 20 reactions</div></td>
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                                      </tr>
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                                      <tr>
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                                        <td width="20%" valign="top" class="style8"><p align="center" class="style8">PlasmidicDNA</p></td>
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                                        <td width="51%" valign="top" class="style8"><p align="center" class="style8">3 &mu;l</p></td>
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                                        <td width="29%" valign="top" class="style8"><p align="center" class="style8">60 &mu;l</p></td>
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                                      </tr>
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                                      <tr>
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                                        <td valign="top" class="style8"><p align="center" class="style8">Buffer 2</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">1 &mu;l</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">20 &mu;l</p></td>
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                                      </tr>
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                                      <tr>
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                                        <td valign="top" class="style8"><p align="center" class="style8">BSA</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">0.5&mu;l</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">10 &mu;l</p></td>
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                                      </tr>
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                                      <tr>
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                                        <td valign="top" class="style8"><p align="center" class="style8">EcoRI</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">0.5 &mu;l / 5 &mu;ll</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">10 &mu;l / 100 &mu;l</p></td>
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                                      </tr>
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                                      <tr>
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                                        <td valign="top" class="style8"><p align="center" class="style8">H<span class="style12">2</span>O</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">5 &mu;l / 10 &mu;l&nbsp;&nbsp; vf</p></td>
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                                        <td valign="top" class="style8"><p align="center" class="style8">100 &mu;l / 200 &mu;l&nbsp;&nbsp; vf</p></td>
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                                      </tr>
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                                    </table>
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                                    <table width="147" height="97" border="0">
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                                      <tr>
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                                        <td height="75">&nbsp;</td>
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                                      </tr>
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                                    </table>
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                                    <p>&nbsp;</p>
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                                    <p>&nbsp;</p>
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                                    <p>&nbsp;</p>
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                                    <p>&nbsp;</p>
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                                    <p>&nbsp;</p>
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                                    <p>Electrophoresis gel load:</p>
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                                    <ul>
 +
                                      <li>3 &mu;l of colorant</li>
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                                      <li>10 &mu;l of digestion</li>
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                                      <li>3 &mu;l of molecular weight marker</li>
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                                    </ul>
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                                    <p>&nbsp;</p>
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                                    <p>Load order.</p>
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                                    <p>Well&nbsp;&nbsp;&nbsp;&nbsp; Plasmid&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Well &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Plasmid</p>
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                                    <p>2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 16,2&rdquo;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 13&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3.3<br>
 +
3&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 4:23  L2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 14&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 22.1<br>
 +
4&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 15:3  L2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 15&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 19.2*<br>
 +
5&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 18.2*  &nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 16&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 24.2&rdquo;<br>
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6&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2.3</p>
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                                    <p>Well&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Well<br>
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  1&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 1kb  2 &mu;l&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 12&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2.3<br>
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  2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 16.2&rdquo;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 13&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 14.2<br>
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  3&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 4:23  L2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 14&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3.3<br>
 +
  4&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 15:3  L2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 15&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 22.1<br>
 +
  5&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 18.2&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 15&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 19.1*<br>
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  6&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 15.13</p>
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                                    <p align="left">&nbsp;</p>
 +
                                    <p>Horizontal Gene Transfer (HGT).<br>
 +
                                      Acording to the model proposed to Arturo,  we made a search for the candidate bioparts to make their elusion and  transformation, which got in the next way:</p>
 +
                                    <p>2 Constructions:</p>
 +
                                    <p><br>
 +
                                      Construction 1:<br>
 +
                                      Biopart BBa_K145201</p>
 +
                                    <p>J23116 + B0034 + C0040 + B0010 + B0012</p>
 +
                                    <p>Construction 2:<br>
 +
                                      The first option for the construction 2 is  made up from two devices: BBa_J01101 and Bba_I763011. The second one consists  in asking Boston for the construction BBa_I763027. We will tell Arturo about  this posibility.<br>
 +
                                      First option:<br>
 +
                                      Device 1.<br>
 +
                                      B0040 + B0034 + C0051 + B0010 + B0012<br>
 +
                                      Device 2.<br>
 +
                                      B0051 + B0034 + J04031</p>
 +
                                    <p align="left">&nbsp;</p>
 +
                                    <p align="left" class="style8">LOCATION</p>
 +
                                    <table width="379" border="1" cellpadding="0"  cellspacing="0">
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                                      <tr>
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                                        <td width="115" valign="top"><p>&nbsp; </p></td>
 +
                                        <td width="99" valign="top" ><p><strong>Plate</strong></p></td>
 +
                                        <td width="51" valign="top" ><p><strong>Well</strong></p></td>
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                                        <td width="104" valign="top" ><p><strong>Plasmid</strong></p></td>
 +
                                      </tr>
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                                      <tr>
 +
                                        <td width="115" valign="top"><p>Device 1    (piece1)</p></td>
 +
                                        <td width="99" valign="top" ><p>2009    kitplate1</p></td>
 +
                                        <td width="51" valign="top" ><p>11H</p></td>
 +
                                        <td width="104" valign="top" ><p>PSB1A2</p></td>
 +
                                      </tr>
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                                      <tr>
 +
                                        <td width="115" valign="top"><p>Device 2    (piece 2)</p></td>
 +
                                        <td width="99" valign="top" ><p>2009    kitplate1</p></td>
 +
                                        <td width="51" valign="top" ><p>15L</p></td>
 +
                                        <td width="104" valign="top" ><p>PSB1A2</p></td>
 +
                                      </tr>
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                                    </table>
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                                    <p align="left" class="style8">&nbsp;</p>
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                                    <p align="left" >&nbsp;</p>
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                                    <p align="left" >&nbsp;</p>
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                                    <table width="100%" height="108" border="0" align="left">
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                                      <tr>
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Revision as of 07:34, 14 October 2009



UNAMFCIENCIASCINVESTAV
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August 10th, 2009

Calculus for Mix; plasmid linearization.

For 20 reactions.

 

 
Mix
For every Eppendorf tube
For 20 reactions

PlasmidicDNA

3 μl

60 μl

Buffer 2

1 μl

20 μl

BSA

0.5μl

10 μl

EcoRI

0.5 μl / 5 μll

10 μl / 100 μl

H2O

5 μl / 10 μl   vf

100 μl / 200 μl   vf
 

 

 

 

 

 

Electrophoresis gel load:

  • 3 μl of colorant
  • 10 μl of digestion
  • 3 μl of molecular weight marker

 

Load order.

Well     Plasmid            Well             Plasmid

2          16,2”               13            3.3
3          4:23 L2            14            22.1
4          15:3 L2            15            19.2*
5          18.2*               16            24.2”
6          2.3

Well                             Well
1          1kb 2 μl         12            2.3
2          16.2”             13            14.2
3          4:23 L2          14            3.3
4          15:3 L2          15            22.1
5          18.2              15            19.1*
6          15.13

 

Horizontal Gene Transfer (HGT).
Acording to the model proposed to Arturo, we made a search for the candidate bioparts to make their elusion and transformation, which got in the next way:

2 Constructions:


Construction 1:
Biopart BBa_K145201

J23116 + B0034 + C0040 + B0010 + B0012

Construction 2:
The first option for the construction 2 is made up from two devices: BBa_J01101 and Bba_I763011. The second one consists in asking Boston for the construction BBa_I763027. We will tell Arturo about this posibility.
First option:
Device 1.
B0040 + B0034 + C0051 + B0010 + B0012
Device 2.
B0051 + B0034 + J04031

 

LOCATION

 

Plate

Well

Plasmid

Device 1 (piece1)

2009 kitplate1

11H

PSB1A2

Device 2 (piece 2)

2009 kitplate1

15L

PSB1A2

 

 

 

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