Team:EPF-Lausanne/Tools/Gibson assembly

From 2011.igem.org

(Difference between revisions)
(Step-by-step)
(Step-by-step)
Line 30: Line 30:
# '''Transforming cells:''' After the reaction, simply use a few microliters of it to transform cells.
# '''Transforming cells:''' After the reaction, simply use a few microliters of it to transform cells.
# '''Negative control:''' For the negative control, make a Gibson reaction on just the plasmid backbone DNA and transform it. This will show you how much of the backbone closes itself up in your actual Gibson reaction.
# '''Negative control:''' For the negative control, make a Gibson reaction on just the plasmid backbone DNA and transform it. This will show you how much of the backbone closes itself up in your actual Gibson reaction.
 +
 +
=== Preparing the parts ===
=== Preparing the parts ===
Line 37: Line 39:
Right overhang (red):
Right overhang (red):
-
* The '''backbone''' has a dark red extension, whose sequence is the last 20bp of the RFP fragment
+
* The '''backbone''' has a dark red extension, whose sequence is the '''last 20bp of the RFP fragment'''
-
* The RFP fragment has an orange extension, whose sequence is the first 20bp of the backbone
+
* The '''RFP fragment''' has an orange extension, whose sequence is the '''first 20bp of the backbone
 +
'''
 +
[[File:EPFL_Gibson_overhangs.jpg]]
== Advantages ==
== Advantages ==

Revision as of 07:53, 12 September 2011