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Degrade DNA to Get Deoxyribose for GC/MS Analysis

  1. Dissolve 100ug DNA in 200ul distilled water.
  2. Add MgCL2 and sodium acetate to the final concentration of 15mM and 10mM, respectively.
  3. Incubate with 2 units DNAse I at room temperature for 15 minutes.
  4. Add ammonium bicarbonate to the final concentration of 100mM, and then incubate with 2mU            phosphodiesterase at 37oC in water bath for 2 hours.
  5. Add 1 unit of alkaline phosphatase and incubated at 37oC water bath for another 1 hour.
  6. Air dry the solution with degraded DNA. Derivalize produced deoxyribose to its aldonitrile acetate form as usual for analysis on GC/MS spectrometer.

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