Team:Macquarie Australia/Protocols/Ligation

From 2013.igem.org

(Difference between revisions)
Line 12: Line 12:
</style>
</style>
 +
 +
A reaction mixture was prepared containing,
<div class="datagrid"><table>
<div class="datagrid"><table>
Line 28: Line 30:
</head>
</head>
</html>
</html>
-
 
-
 
-
 
-
A reaction mixture was prepared containing,
 
-
Element Volume
 
-
Upstream Digestion part 2 µL
 
-
Downstream Digestion part 2 µL
 
-
Destination Plasmid 1 µL
 
-
10X T4 DNA ligase buffer 2 µL
 
-
T4 DNA ligase 1 µL
 
-
H2O 12 µL
 
-
 
   
   
<br><br>
<br><br>

Revision as of 05:16, 29 August 2013


Ligation Procedure

A reaction mixture was prepared containing,

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



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

4 µL of the ligation product was transformed into 100 µL of competent E. coli, the transformants were then incubated for one hour.

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