DNA Degradase Plus

E2020 / E2021


DNA Degradase Plus

E2020 / E2021

Cat # Name Size Price Quantity
E2020 DNA Degradase Plus 250 U $136.00
+ -
E2021 DNA Degradase Plus 1000 U $435.00
+ -

Documents


Data Sheet: PDF   |   Protocol: PDF   |   SDS (MSDS): PDF   |  
Complete digestion of DNA into individual nucleosides.

Highlights


  • Quick and simple procedure for generating single nucleosides from DNA for quantitative analysis via LC/MS
  • Convienent 2 hour, single enzyme digest v. conventional 6-16 hr multi-step enzyme digestion protocol

Description


DNA Degradase Plus from Zymo Research is a nuclease mix that quickly and efficiently degrades DNA to its individual nucleoside components. Since nucleosides lack negatively charged phosphate, DNA Degradase Plus is ideal for whole-genome DNA methylation analysis by LC/MS. Digestion with the enzyme is performed via a one-step procedure that is faster and simpler than other available methods.

Technical Specifications


Assay Condition DNA Degradase Plus in 1X DNA Degradase Reaction Buffer. Incubate reaction mixtures at 37C for ≥1 hour
Concentration 5 U/µl
Enzyme Inactivation Heat inactivate enzyme at 70C for 20 minutes
Storage Store at -20C for up to 12 months. Avoid repeated freeze/thawing of reagents. Prolonged storage is at ≤ -70C.
Unit Definition 5 U of enzyme will digest 1 µg of genomic DNA in 25 µl reaction volume at 37C for ≥ 1 hr

Product FAQ


Q1: Can I visualize the treated samples/ do I need to purify after Degradase treatment?

Q2: Can I let the reaction go longer than stated in the protocol?

Q3: Can I add excess enzyme to the reaction?

Q4: Does this work with RNA/ ssDNA?

Q5: Can I scale up/ down the reaction volumes? How do I do that?

Citations


The authors used DNA Degradase Plus to determine total methylation in genomic DNA from developing mouse embryonic tissue by mass spectrometry.

Oda M, Oxley D, Dean W, Reik W. 2013. Regulation of Lineage Specific DNA Hypomethylation in Mouse Trophectoderm. Plos One DOI: 10.1371/journal.pone.0068846.

The authors sought to quantify levels of 5-mC in a number of yeast species using DNA Degradase Plus and LC-MS/MS.

Capuano F, Mülleder M, Kok R, Blom HJ, Ralser M. 2014. Cytosine DNA Methylation Is Found in Drosophila melanogaster but Absent in Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Other Yeast Species. Anal. Chem. 86(8): 3697–3702.

DNA Degradase Plus was used to quantify the levels of 5-hmC and 5-mC in genomic DNA of tumor cells or cultured cell lines using a sensitive liquid chromatography-tandem quadrupole mass spectrometric method.

Tsuji M, Matsunaga H, Jinno D, Tsukamoto H, Suzuki N, Tomioka Y. 2014. A validated quantitative liquid chromatography-tandem quadrupole mass spectrometry method for monitoring isotopologues to evaluate global modified cytosine ratios in genomic DNA. J Chro

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