From 2007.igem.org

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For 20uL reation volume

Reaction Mixture

• 0.5uL Enzyme 1
• 0.5uL Enzyme 2
  • Add enzymes last
• 2uL appropriate 10x buffer (see table on fridge)
• 2uL 10x BSA
• 10uL MilliQ
• 5uL DNA
  • If performing the same digest for multiple DNA samples make up mastermix of buffer enzyme soultion and make 15uL aliquots to which to add the DNA
  • Volume DNA may vary depending on concentration, vary water volume to achieve 20uL reaction volume.

1. Incubate for 1-3 hrs at 37 degrees.
2. Stop reaction with addition of 5uL 6x DNA loading dye.
3. Can be stored at -20 or run on gel immediately.