Team:Washington/GENERAL LIGATION PROTOCOL

From 2013.igem.org

(Difference between revisions)
(→')
(→')
 
(4 intermediate revisions not shown)
Line 1: Line 1:
-
==''''''==
+
{{:Team:Washington/WA_Template}}
-
<html>
 
 +
 +
<html>
 +
<blockquote>
<header>
<header>
<b>General Ligation Protocol</b>
<b>General Ligation Protocol</b>
Line 27: Line 29:
65C for 10 minutes (or 80C)<br>
65C for 10 minutes (or 80C)<br>
<br>
<br>
-
 
-
 
-
<a href = "https://www.neb.com/protocols/1/01/01/dna-ligation-with-t4-dna-ligase-m0202/">DNA ligation with T4 DNA Ligase</a><br>
 
<br>
<br>

Latest revision as of 04:32, 27 September 2013


General Ligation Protocol


Reaction Mixture: 7uL insert (1:3 to 1:7 ratio of vector:insert) & (2:6) also works well
1uL vector
1uL T4 ligase buffer
1uL T4 ligase
10ul total

Incubate at 16C overnight, or room temperature for 30 minutes to 1 hour.

Heat inactivation after ligation, before transformation can increase transformation efficiency.
65C for 10 minutes (or 80C)



Retrieved from "http://2013.igem.org/Team:Washington/GENERAL_LIGATION_PROTOCOL"