IGEM:UC Berkeley/2006/Analitic digestion

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  1. Use Ape to determine which enzymes to cut product with. Cuts should yield bands that are easily distinguishable on a gel.
  2. To digest make a cocktail of NEB2, Enzyme, H2O, and DNA product for a total volume of 10uL.
    • If DNA product is high copy then use only 2uL of product 1uL of NEB2 and adjust the H2O and enzyme accordingly to make 10uL total:
        • 2 uL miniprep/DNA product
        • 1 uL NEB2
        • .25 uL enzyme 1
        • .25 uL enzyme 2
        • 1 uL BSA
        • 5.5 uL water
    • If DNA product is low copy then use 4uL of product and 1uL of NEB2 adjusting the H2O and enzyme accordingly:
        • 4 uL miniprep/DNA product
        • 1 uL NEB2
        • .25 uL enzyme 1
        • .25 uL enzyme 2
        • 4.5 uL water
  3. Incubate for ~30min. and then run on gel.
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