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Team:Valencia/Dna Digestion
From 2012.igem.org
DNA Digestion
Reagents and Materials
For a DNA digestion of total volume V (in μL):
- X µl 1-520 ng.
- 5 µl NEB2 buffer.
- 0.5 µl 100x BSA.
- 1 μL BB enzyme 1.
- 1 μL BB enzyme 2.
- (42.5-x) µl distilled water.
Protocol
- Find appropriate enzymes(s), associated buffer, reaction temperature from NEB.
- Calculate the total volume of the reaction by taking the DNA volume + 1(2) μL for a single (double) digestion reaction. Divide this number by 0.8 to get the total reaction volume, V.
- Calculate the volume of 10mg/ml BSA and 10X digestion buffer, which are both 0.1V.
- Add the reaction components together in order listed above. Mix gently and spin.
- Incubate at reaction temperature for 3 hr to overnight.
