Team:Macquarie Australia/Protocols/Ligation

From 2013.igem.org

(Difference between revisions)
Line 12: Line 12:
</style>
</style>
 +
<br>
-
A reaction mixture was prepared containing,
+
<b>Prepare</b> the following reaction mixture
<div class="datagrid"><table>
<div class="datagrid"><table>
-
<table align="center"> <!-- Added by me -->
+
<!--      <table align="center"> Added by me -->
<thead><tr><th>Element </th><th>Volume</th></tr></thead>
<thead><tr><th>Element </th><th>Volume</th></tr></thead>
<tbody><tr><td>Upstream Digestion part </td><td>2 µL</td></tr>
<tbody><tr><td>Upstream Digestion part </td><td>2 µL</td></tr>
Line 31: Line 32:
</html>
</html>
   
   
 +
Incubate each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.
<br><br>
<br><br>
-
We incubated each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.
+
transform 4 µL of the ligation product into 100 µL of competent E. coli, then incubate the transformants for one hour.
-
<br><br>
+
-
4 µL of the ligation product was transformed into 100 µL of competent E. coli, the transformants were then incubated for one hour.
+

Revision as of 07:05, 13 September 2013


Ligation Procedure


Prepare the following reaction mixture

Element Volume
Upstream Digestion part 2 µL
Downstream Digestion part2 µL
Destination Plasmid1 µL
10X T4 DNA ligase buffer2 µL
T4 DNA ligase1 µL
H2O12 µL

Incubate each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.

transform 4 µL of the ligation product into 100 µL of competent E. coli, then incubate the transformants for one hour.

Retrieved from "http://2013.igem.org/Team:Macquarie_Australia/Protocols/Ligation"