Difference between revisions of "Team:EPF Lausanne/Notebook/Protocols"

Line 193: Line 193:
 
<p> • Tryptophan</p>
 
<p> • Tryptophan</p>
 
<h2>Procedure</h2>
 
<h2>Procedure</h2>
<table width="100%">
+
<table width=”100%>
<thread>
+
 
<tr>
 
<tr>
 
<th>Stock concentration</th>
 
<th>Stock concentration</th>
Line 200: Line 199:
 
<th>Total quantity for 50 mL</th>
 
<th>Total quantity for 50 mL</th>
 
</tr>
 
</tr>
</thread>
 
<tbody>
 
 
<tr>
 
<tr>
<th>100 mM Histidine-Hcl (209 g/mol)</th>
+
<td>100 mM Histidine-Hcl (209 g/mol)</td>
<th> 20.9 g/L</th>
+
<td> 20.9 g/L</td>
<th> 0.418 g</th>
+
<td> 0.418 g</td>
 
</tr>
 
</tr>
 
<tr>
 
<tr>
<th>20 mM Uracil (112 g/mol)</th>
+
<td>20 mM Uracil (112 g/mol)</td>
<th> 2.24 g/L</th>
+
<td> 2.24 g/L</td>
<th> 0.0448 g</th>
+
<td> 0.0448 g</td>
 
</tr>
 
</tr>
 
<tr>
 
<tr>
<th>100 mM Leucine (131 g/mol)</th>
+
<td>100 mM Leucine (131 g/mol)</td>
<th> 13.1 g/L</th>
+
<td> 13.1 g/L</td>
<th> 0.262 g</th>
+
<td> 0.262 g</td>
 
</tr>
 
</tr>
 
<tr>
 
<tr>
<th>40 mM Tryptophan (204 g/mol)</th>
+
<td>40 mM Tryptophan (204 g/mol)</td>
<th> 8.16 g/L</th>
+
<td> 8.16 g/L</td>
<th> 0.1632 g</th>
+
<td> 0.1632 g</td>
 
</tr>
 
</tr>
</tbody>
 
 
</table>
 
</table>
 
<p> • Filter and sterilize solutions</p>
 
<p> • Filter and sterilize solutions</p>

Revision as of 13:27, 21 July 2015

Protocols

Agarose Gel

Materials

• 1X TAE

• Agarose

• Gel Red

• DNA samples

• 6X loading dye

• Nuclease free water

Procedure

Prepare 1.2% agarose gel for small fragments and 3% agarose gel for large fragments

• Mix 50 mL 1X TAE and 0.6 g (1.2%) or 1.5 g (3%) agarose

• Melt in microwave until agarose has melted (about 50 seconds)

• Add 1.3 μL (1.2%) or 1.5 μL (3%) Gel Red

• Pour solution into agarose gel mold with comb

• Let set for 20 minutes or until solid

• Place gel in 1X TAE and remove comb

• Load samples of 200 ng (or 2 μL) DNA mixed with 2 μL 6X loading dye and nuclease free water up to 12 μL

• Run gel at 100-120 Volts for 40-50 minutes (1.2%) or 80 Volts for 2 hours (3%)

• Take a picture of the gel at the UV detector

Amino acid solution

Materials

• Histidine-Hcl

• Uracil

• Leucine

• Tryptophan

Procedure

Stock concentration Final concentration Total quantity for 50 mL
100 mM Histidine-Hcl (209 g/mol) 20.9 g/L 0.418 g
20 mM Uracil (112 g/mol) 2.24 g/L 0.0448 g
100 mM Leucine (131 g/mol) 13.1 g/L 0.262 g
40 mM Tryptophan (204 g/mol) 8.16 g/L 0.1632 g

• Filter and sterilize solutions

•Add 8 mL per liter of selective medium or spread 500 μL on a selective plate

Section 3

Try to scroll this section and look at the navigation list while scrolling!

Section 4-1

Try to scroll this section and look at the navigation list while scrolling!

Section 4-2

Try to scroll this section and look at the navigation list while scrolling!

Still under construction