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proteinase K problem


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#1 preeti_RNA

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Posted 13 January 2010 - 06:39 AM

Hi everyone,

I am facing problem of DNA loss during proteinaseK treatment of genomic DNA of plant(60oC, 3h). I have tried diff. conc. of proteinase K but its degrading my DNA. All of my colleagues are using proteinase K for other organism on which its working fine so no question of having contamination. If anyone can please suggest me how to get rid of this problem asap

Thanks

Preeti

#2 HomeBrew

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Posted 13 January 2010 - 03:08 PM

How did you conclude that the PK is degrading your DNA? Did you look at it on a gel? What happens if you add just sterile water (or whatever your PK diluent is) to your DNA (use the same volume of water that you would add of PK solution) and incubate it at 60 C for 3 hours? Is this mock sample degraded as well?

Is your proteinase K certified as DNAse/RNase free?




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