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DNA stability in STE with proteinase K


Best Answer Trof, 18 June 2014 - 12:17 PM

Yes, it's pretty stable in STE at -20. Sometimes we had them there for months, maybe even years (but I'm not sure about quality of that). However, tail DNA is always not exactly a good quality, full of PCR inhibitors. If you need it for PCR, dilute to low concentrations like 10-20 ng/ul and use more PCR cycles.

 

Lysates stored frozen are generaly more stable than original material, that's particulary important for RNA isolation.

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#1 SG181

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Posted 18 June 2014 - 09:45 AM

I'm currently working on genotyping mouse tail snips, however I won't be able to do the phenol-chloroform extractions for several days. If I performed the digest of the tail snips in STE (50 mM Tris, 100 mM NaCl, 1mM EDTA, 1%SDS) with proteinase K, would the samples be stable for up to a week if I stored them at -20°C once I finished the digest? Or should I just wait and do the digest the day before I can do the phenol-chloroform extractions so that I avoid potential degradation of the DNA?



#2 Trof

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Posted 18 June 2014 - 12:17 PM   Best Answer

Yes, it's pretty stable in STE at -20. Sometimes we had them there for months, maybe even years (but I'm not sure about quality of that). However, tail DNA is always not exactly a good quality, full of PCR inhibitors. If you need it for PCR, dilute to low concentrations like 10-20 ng/ul and use more PCR cycles.

 

Lysates stored frozen are generaly more stable than original material, that's particulary important for RNA isolation.


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