my DNA is degrading..

[noa]

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???

[admin]

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