Team:Newcastle/Ligation

From 2010.igem.org

(Difference between revisions)
(Procedures)
Line 6: Line 6:
A ligation calcultion can be used however a 1:3 and a 1:5 ratio tends to be used instead.  
A ligation calcultion can be used however a 1:3 and a 1:5 ratio tends to be used instead.  
-
=Procedures=
+
==Procedures==
Set up the ligation mix according:
Set up the ligation mix according:

Revision as of 15:12, 29 July 2010

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Ligation

For standard ligation 100-200ng of vector DNA is required - can be checked using nanodrop

A ligation calcultion can be used however a 1:3 and a 1:5 ratio tends to be used instead.

Procedures

Set up the ligation mix according:

Reagents 1:3(μl) 1:5(μl) Vector(μl)
V 1 1 1
I 3 5
10*BUFFER 1 1 1
Ligase T4 1 1 1
H2O 4 2 7
Total Volume 10.0 10.0 10.0

For sticky end ligations incubate on the bench of 3-4 hrs or overnight

Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon