Promega Quick Ligation
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1.CIP treatment
DNA: 15μL
10x CIP: 0.43μL
10x Buffer: 1.72μL
Total: 17.2μL
2. Assemble the following reaction in a sterile microcentrifuge tube:
vector DNA 100ng
insert DNA 33ng
2X Rapid Ligation Buffer 5μl
T4 DNA Ligase (Weiss units) 3u
Nuclease-Free Water to final volume of 10μl
2. Incubate the reaction at room temperature for 5 minutes for cohesive-ended ligations, or 15 minutes for blunt-ended ligations.rature for 5 minutes for cohesive-ended ligations, or 15 minutes for blunt-ended ligations.
