TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol covers the inoculation, cultivation, IPTG-induction and plate reader measurement of <em>E. coli</em> BL21 strains carrying constructs with various toehold switch devices from Green <em>et al</em>., 2014.</p>

[BEFORE_START]
<p> </p>
<table style="height: 184px; width: 265px;">
<tbody>
<tr>
<td style="width: 160.71875px;"><strong>Materials/ reagents</strong></td>
<td style="width: 88.296875px;"><strong># per group</strong></td>
</tr>
<tr>
<td style="width: 160.71875px;">Culture tubes</td>
<td style="width: 88.296875px;">3x</td>
</tr>
<tr>
<td style="width: 160.71875px;">Erlenmeyer flasks, 100mL</td>
<td style="width: 88.296875px;">3x</td>
</tr>
<tr>
<td style="width: 160.71875px;">12-well plate</td>
<td style="width: 88.296875px;">1</td>
</tr>
<tr>
<td style="width: 160.71875px;">Cuvettes</td>
<td style="width: 88.296875px;">as needed</td>
</tr>
<tr>
<td style="width: 160.71875px;">LB, sterile</td>
<td style="width: 88.296875px;">as needed </td>
</tr>
<tr>
<td style="width: 160.71875px;">H<sub>2</sub>O, sterile</td>
<td style="width: 88.296875px;">as needed </td>
</tr>
</tbody>
</table>
<p> </p>
<table style="height: 127px; width: 178.046875px;">
<tbody>
<tr>
<td style="width: 81px;"><strong>Stock solutions</strong></td>
<td style="width: 96.046875px;"><strong>Stock concentration</strong></td>
</tr>
<tr>
<td style="width: 81px;">IPTG</td>
<td style="width: 96.046875px;">100 mM</td>
</tr>
<tr>
<td style="width: 81px;">Kanamycin</td>
<td style="width: 96.046875px;">100 mg/mL</td>
</tr>
<tr>
<td style="width: 81px;">Ampicillin</td>
<td style="width: 96.046875px;">25 mg/mL</td>
</tr>
</tbody>
</table>

[GUIDELINES]
<p>The class will divide into four groups; each group will investigate one separate RNA device from Green <em>et al</em>. </p>
<p>Each device consists of two plasmids - a switch plasmid containing GFPmut3b, cis-repressed by a synthetic regulatory 5'UTR (untranslated region), denoted by H-xx-GFP, and a trigger plasmid containing a small regulatory RNA, denoted by T-xx, which binds to the switch sequence and switches on translation of GFP.</p>
<p>The following table shows the strains used by each group:</p>
<p> </p>
<table style="height: 148px; width: 550px;">
<tbody>
<tr style="height: 18px;">
<td style="width: 130px; height: 18px;"> </td>
<td style="width: 130px; height: 18px;">Toehold switch device</td>
<td style="width: 130px; height: 18px;">Switch only control</td>
<td style="width: 154.328125px; height: 18px;">non-specific trigger control</td>
<td style="width: 10px; height: 18px;"> </td>
</tr>
<tr style="height: 24px;">
<td style="width: 130px; height: 24px;">
<p>Group 1</p>
</td>
<td style="width: 130px; height: 24px;">  T56, H56-GFP</td>
<td style="width: 130px; height: 24px;"> H56-GFP</td>
<td style="width: 154.328125px; height: 24px;">  T117, H56-GFP</td>
<td style="width: 10px; height: 24px;"> </td>
</tr>
<tr style="height: 17px;">
<td style="width: 130px; height: 17px;">Group 2</td>
<td style="width: 130px; height: 17px;">  T69, H69-GFP</td>
<td style="width: 130px; height: 17px;"> H69-GFP</td>
<td style="width: 154.328125px; height: 17px;">   T56, H69-GFP</td>
<td style="width: 10px; height: 17px;"> </td>
</tr>
<tr style="height: 15px;">
<td style="width: 130px; height: 15px;">Group 3</td>
<td style="width: 130px; height: 15px;">  T96, H96-GFP</td>
<td style="width: 130px; height: 15px;"> H96-GFP</td>
<td style="width: 154.328125px; height: 15px;">   T69, H96-GFP</td>
<td style="width: 10px; height: 15px;"> </td>
</tr>
<tr style="height: 20px;">
<td style="width: 130px; height: 20px;">Group 4</td>
<td style="width: 130px; height: 20px;">  T117, H117-GFP</td>
<td style="width: 130px; height: 20px;"> H117-GFP</td>
<td style="width: 154.328125px; height: 20px;">   T96, H117-GFP</td>
<td style="width: 10px; height: 20px;"> </td>
</tr>
</tbody>
</table>
<p> </p>

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