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Team:Hannover/Protocols/Cloning/Ligation
From 2014.igem.org
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<p class="text">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. </p> | <p class="text">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. </p> | ||
| - | <h2>Table 1: Reaction | + | <h2>Table 1: Reaction Mixes and Temperature Programs for the Ligation.</h2> |
<table align="line" width="800px"><tr><td><table> | <table align="line" width="800px"><tr><td><table> | ||
<tr><td><b>Volume [μl]</b></td><td><b>Compounds of the digest</b></td></tr> | <tr><td><b>Volume [μl]</b></td><td><b>Compounds of the digest</b></td></tr> | ||
Revision as of 19:15, 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.
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