"
Team:Hannover/Protocols/Cloning/Ligation
From 2014.igem.org
(Difference between revisions)
(Created page with "{{Template:Team:Hannover/igem.css}} {{Template:Team:Hannover/main.css}} {{Template:Team:Hannover/main.tmpl}} {{Template:Team:Hannover/newmenu.tmpl}} <html> <body> <h1><a href="ht...") |
|||
| Line 14: | Line 14: | ||
<tr><td>2.00</td><td>10 x T4 DNA Ligase buffer </td></tr> | <tr><td>2.00</td><td>10 x T4 DNA Ligase buffer </td></tr> | ||
<tr><td>2.00</td><td>10 mM µl-1 ATP</td></tr> | <tr><td>2.00</td><td>10 mM µl-1 ATP</td></tr> | ||
| - | <tr><td>1.00</td><td>1 | + | <tr><td>1.00</td><td>1 T4 DNA Ligase</td></tr> |
<tr><td> -</td><td>20-100 ng linear vector DNA</td></tr> | <tr><td> -</td><td>20-100 ng linear vector DNA</td></tr> | ||
<tr><td> -</td><td>100-500 ng insert DNA</td></tr> | <tr><td> -</td><td>100-500 ng insert DNA</td></tr> | ||
Revision as of 14:08, 15 October 2014
Protocols / Ligation
After restriction enzyme cleavage, compatible ends of two fragments can hybridize, but are not stable due to their missing phosphodiester bonds. To fix those nicks in the double-stranded DNA, a ligase from T4 phages (Thermo Scientific) was utilized. The ligation reaction mix is shown in the following table.
Table 1: Reaction mixes and temperature programs for the ligation.
|
| |||||||||||||||||||||||||||
