Melbourne/Diagnostic Digest

From 2007.igem.org

(Difference between revisions)
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==For 20uL reation volume==
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===For 20uL reation volume===
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===Reaction Mixture===
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====Reaction Mixture====
*0.5uL Enzyme 1
*0.5uL Enzyme 1
*0.5uL Enzyme 2
*0.5uL Enzyme 2
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**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
**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.
**Volume DNA may vary depending on concentration, vary water volume to achieve 20uL reaction volume.
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#Incubate for 1-3 hrs at 37 degrees.
#Incubate for 1-3 hrs at 37 degrees.
#Stop reaction with addition of 5uL 6x DNA loading dye.
#Stop reaction with addition of 5uL 6x DNA loading dye.
#Can be stored at -20 or run on gel immediately.
#Can be stored at -20 or run on gel immediately.

Revision as of 06:23, 8 July 2007

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.