IGEM:Hong Kong UST 2014/2009/Notebook/ATP Concentration Effect to Ligation Speed/2014/06/05
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Thursday, 5th of June 2014
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11. Making the ATP Stock
Make 10 mM ATP stock for 1 ml by doing serial dilution.
The ATP solution then sterilized with filter sterilization
Make the recipe for ligation. Each person in the team will use 4 micro centrifuge tube to create the control, 1mM ATP, 2mM ATP, and 4 mM ATP. The amount of insert and backbone added varies due to the different amount of concentration of nucleic acid in each set.
The ratio of insert to backbone must be 3:1. And since the length of the insert and backbone is 1 kb to 2 kb respectively, the amount of insert must be 30 ng and backbone must be 20 ng. ( total amount of both insert and backbone is ~50 ng)
Incubate in room temperature for 50 minutes