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DNA ligation: Difference between revisions
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#Add appropriate amount of insert to the tube. | #Add appropriate amount of insert to the tube. | ||
#Add appropriate amount of vector to the tube. | #Add appropriate amount of vector to the tube. | ||
#Add 0.5 μL ligase. <br>Vortex ligase before pipetting to ensure that it is well-mixed. <br>Also, the ligase, like most enzymes, is in some percentage of glycerol which tends to stick to the sides of your tip. To ensure you add only | #Add 0.5 μL ligase. <br>Vortex ligase before pipetting to ensure that it is well-mixed. <br>Also, the ligase, like most enzymes, is in some percentage of glycerol which tends to stick to the sides of your tip. To ensure you add only 0.5 μL, just touch your tip to the surface of the liquid when pipetting. | ||
#Let the 10 μL solution sit at 22.5°C for 30 mins | #Let the 10 μL solution sit at 22.5°C for 30 mins | ||
#Denature the ligase at 65°C for 10min | #Denature the ligase at 65°C for 10min | ||
