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Team:Groningen/Template/MODULE/Notebook/secretion/week9
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| - | September | + | September 29 - October 5 |
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<div class="text"> | <div class="text"> | ||
| - | <b>goal:</b> | + | <b>goal:</b> Obtaining pLASs with 3-a assembly |
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<div class="text"> | <div class="text"> | ||
| - | + | Extra amounts of <i>dspB</i>, <i>dspB+HIS</i>, <i>aiiA</i>, <i>aiiA+HIS</i>,<i>aiiA+HIS</i>, <i>pLAS</i>, RBS, double terminator, pSB2k3, pSB1C3 and pSB1A3 were miniprepped to be used for the assembly | |
</div> | </div> | ||
<div class="hspacer"> </div> | <div class="hspacer"> </div> | ||
<!-- PARAGRAPH SNIPPET END--> | <!-- PARAGRAPH SNIPPET END--> | ||
| + | |||
| + | <!-- PARAGRAPH SNIPPET START--> | ||
| + | <div class="text"> | ||
| + | |||
| + | 1 ug of <i>aiiA</i>, <i>dspB</i>, <i>dspB+HIS</i> and <i>aiiA+HIS</i> with XbaI and PstI and 1 ug of RBS was cut with EcoRI and SpeI | ||
| + | |||
| + | </div> | ||
| + | <div class="hspacer"> </div> | ||
| + | <!-- PARAGRAPH SNIPPET END--> | ||
| + | |||
| + | <!-- PARAGRAPH SNIPPET START--> | ||
| + | <div class="text"> | ||
| + | |||
| + | These were eventually ligated for 3 hours into pSB2K3 and transformed into <i>E. coli</i> NEB cells: | ||
| + | </div> | ||
| + | <div class="hspacer"> </div> | ||
| + | <!-- PARAGRAPH SNIPPET END--> | ||
| + | |||
| + | <!-- TABLE SNIPPET OPTIONAL START--> | ||
| + | <table class="table"> | ||
| + | <tr> | ||
| + | <th>Name assembly</th><th>Upstream part</th><th>Downstream part</th><th>Backbone</th> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>A1’</td><td>RBS</td><td><i>aiiA</i></td><td>pSB2K3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>A2’</td><td>RBS</td><td><i>dspB</i></td><td>pSB2K3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>A3’</td><td>RBS</td><td><i>dspB+HIS</i></td><td>pSB2K3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>A4’</td><td>RBS</td><td><i>aiiA+HIS</i></td><td>pSB2K3</td> | ||
| + | </tr> | ||
| + | |||
| + | </table> | ||
| + | <div class="hspacer"> </div> | ||
| + | <!-- TABLE SNIPPET OPTIONAL END--> | ||
| + | |||
| + | |||
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Revision as of 20:48, 17 October 2014
September 29 - October 5
goal: Obtaining pLASs with 3-a assembly
Extra amounts of dspB, dspB+HIS, aiiA, aiiA+HIS,aiiA+HIS, pLAS, RBS, double terminator, pSB2k3, pSB1C3 and pSB1A3 were miniprepped to be used for the assembly
1 ug of aiiA, dspB, dspB+HIS and aiiA+HIS with XbaI and PstI and 1 ug of RBS was cut with EcoRI and SpeI
These were eventually ligated for 3 hours into pSB2K3 and transformed into E. coli NEB cells:
| Name assembly | Upstream part | Downstream part | Backbone |
|---|---|---|---|
| A1’ | RBS | aiiA | pSB2K3 |
| A2’ | RBS | dspB | pSB2K3 |
| A3’ | RBS | dspB+HIS | pSB2K3 |
| A4’ | RBS | aiiA+HIS | pSB2K3 |
