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DNase Treatment

Description
A DNase treatment is conducted to remove genomic DNA from total RNA before reverse transcription.  The Genome Core uses Promega RQ1 RNase-Free DNase.  This step is essential when using Sybr Green RT-PCR chemistry, which will detect all double stranded DNA, including non-specific products.  A DNase treatment is also necessary when using a primer/probe set that does not span an exon/exon boundary.  An Applied Biosystems Assay-on-Demand (AOD) will end in _g1 or _s1 if it will amplify genomic DNA, and a DNase treatment should be strongly considered when using this type of assay.

Science
See information from Promega.

Protocol
Available here.

Pricing
See pricing schedule here.

What to Provide
Please provide the core with the RNA samples for which you would like DNase treatment.  If the RNA samples are to be used in a subsequent reverse transcription and/or expression analysis project, remember to provide enough quantity (volume and/or concentration) to completethe project as requested. One DNase reaction consists of a maximum volume of 8ul of RNA.  The total quantity of RNA is dependent on the size of the real time PCR project. RNA of low concentration may require multiple DNase treatments per sample and will be charged accordingly.

Additional Resources
A searchable database of core facilities at all UCSF campus locations, provided by the Clinical and Translational Science Institute at UCSF, is available here.