Team:Groningen/protocols/Digestion

From 2013.igem.org

(Difference between revisions)
Line 36: Line 36:
<h3>Reaction:</h3>
<h3>Reaction:</h3>
-
{|
+
<table border="1">
-
| Compound || 20 &microl || align="right" | Final concentration
+
  <tr>
-
|-
+
    <th>Compound</th>
-
| Bread  || Pie      || align="right" | 500.00
+
    <th>20&micro;l</th>
-
|-
+
    <th>Final concentration</th>
-
| Butter || Ice cream || align="right" | 1.00
+
  </tr>
-
|}
+
-
<table border = "1">
+
  <tr>
-
<tr><td>Component</td>
+
    <td>MQ water</td>
-
<td>30&#956;l reaction</td>
+
    <td>up to 20&micro;l</td>
-
<td>Final concentration</td>
+
    <td></td>
-
</tr>
+
  </tr>
-
<tr><td>H<sub>2</sub>O</td>
+
 
-
<td>up to 30&#956;l</td>
+
  <tr>
-
<td></td>
+
    <td>10x buffer</td>
-
</tr>
+
    <td>2&micro;l</td>
-
<tr><td>Buffer 10x</td>
+
    <td>1x</td>
-
<td>3&#956;l</td>
+
  </tr>
-
<td>1x</td>
+
 
-
</tr>
+
  <tr>
-
<tr><td>DNA</td>
+
    <td>restriction enzyme</td>
-
<td>4&#956;l</td>
+
    <td>1&micro;l</td>
-
<td>up to 1&#956;g</td>
+
    <td>1U</td>
-
</tr>
+
  </tr>
-
<tr><td>Enzyme/s</td>
+
 
-
<td>1&#956;l</td>
+
  <tr>
-
<td>1U</td>
+
    <td>DNA</td>
-
</tr>
+
    <td>4&micro;l</td>
-
</table>
+
    <td>up to 1&micro;l</td>
 +
  </tr>
 +
 
 +
</table>  
<br>All the reagents are added following the order listed in the table above.
<br>All the reagents are added following the order listed in the table above.

Revision as of 13:57, 27 July 2013

Digestion

Fast digest

Materials:

  • MQ water
  • 1.5ml tubes
  • Fast digestion buffer
  • Restriction enzymes
  • DNA

Reaction:

Compound 20µl Final concentration
MQ water up to 20µl
10x buffer 2µl 1x
restriction enzyme 1µl 1U
DNA 4µl up to 1µl

All the reagents are added following the order listed in the table above.
After the reaction is ready mix the content of the tube and spin it down.
The tubes are incubated for 1h at 37°C.

Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf