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  Answer ID  
3
  Product  
Tech Service: Amplification & Primers
  RT-PCR, cDNA, and RACE Systems
  Date Created  
06/10/1999 04:42 PM
  Date Updated  
07/09/2002 03:24 PM
  Access Level  
Everyone

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  How can I treat my RNA sample before RT-PCR to eliminate DNA contamination?
  Question
  How can I treat my RNA sample before RT-PCR to eliminate DNA contamination?
  Answer
  Combine 1 g total RNA, 1 l 10X DNAse I buffer (200 mM Tris-HCl (pH 8.4),500 mM KCl, 20 mM MgCl2), 1 l DNAse I, Amplification Grade, 1 unit/l and DEPC treated water to 10 l. Incubate for 15 min at room temperature. Inactivate by adding 1 l of 25 mM EDTA and heat for 10 min at 65 C. Note: 1 unit of DNAse I should be enough to treat up to ~10 g of RNA.
 
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