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my DNA is degrading..

my DNA is degrading.. - DNA Extraction Forum

my DNA is degrading.. - DNA Isolation Forum. A forum specifically about questions on DNA extraction and purification.

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Old 12-22-2010, 08:46 AM
noa noa is offline
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Question my DNA is degrading..

Im trying to purify DNA from plasmodium parasite for souther but it keeps degrading! im incubating it with TSE, SDS and proteinase K ON.
does anybody has suggestions for how to prevent DNA from degrading during the purification???
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Old 12-22-2010, 01:29 PM
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Default Re: my DNA is degrading..

I would make sure you have everything on ice just in case, and possibly add a Dnase inhibitor.

Inhibitors of DNase:

  • 2-mercaptoethanol (Laskowski 1971)
  • 2-nitro-5-thiocyanobenzoic acid (Moore 1981)
  • Actin (Mannherz et al. 2008, and Lazarides and Lindberg 1974)
  • Alfatoxin B2a, G2, G2a, and M1 (non-competitive) (Schabort 1970)
  • Ca2+ (Moore 1981)
  • EGTA (Moore 1981) and EDTA (Junowicz and Spencer 1973)
  • Sodium dodecyl sulfate (Liao 1975a)
  • Calf spleen inhibitor protein (Laskowski 1971)
  • Carbodiimide and cholesterol sulfate (Iwamori 2000)
  • Iodoacetate (Moore 1981)

You could also try isolating the nuclear protein extract that would get rid of a lot of DNases.

see this post for more:

DNAse Inhibition for Perfect DNA Isolations

Last edited by admin; 12-22-2010 at 01:35 PM.
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