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Baseline-ZERO DNase
Digest large or small DNA molecules better than standard DNase I.
Key features
Show- Effectively digest large or small dsDNA and ssDNA into mononucleotides
- Digest unwanted dsDNA and ssDNA molecules including small ssDNA such as random hexamers
- Use in sensitive applications requiring reverse transcription where any contaminating DNA is unwanted
Size: 5000 units
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Product information
Baseline-ZERO™ DNase* digests dsDNA and ssDNA into mononucleotides more effectively than the commonly used bovine pancreatic DNase I. Even the small DNA oligonucleotides that remain after treatment with bovine pancreatic DNase I are undetectable by gel electrophoresis following treatment with Baseline-ZERO DNase (Figure. 2). Removal of DNA from RNA preparations is particularly beneficial when RNA in a sample is amplified using a method that involves reverse transcription using random primers, since any contaminating DNA would also be a template for random-primed cDNA synthesis.
Applications
- Removal of genomic DNA from small-sample total RNA preparations for expression analysis (Figure. 1)
- Removal of small DNA oligonucleotides (e.g. random primers)
Figure 1. Real-time PCR of HeLa RNA preparations treated with various DNases. The lower the C T value (intersection of curves with the red line), the greater the amount of residual DNA not digested by the indicated DNase. Thus, Baseline-ZERO™ DNase removed all detectable DNA from the RNA sample. The TaqMan® probe assay amplified a 268-bp fragment of β-actin. Samples were run in duplicate.
References
- Kienzle, N. et al. (1996) BioTechniques 20, 612.
*Patent pending.
Unit Definition: One Molecular Biology Unit (MBU) of Baseline-ZERO™ DNase produces an increase in the A260 of a solution of dsDNA, of 0.001 per minute at 25 °C. Functionally, 1 MBU completely digests 1 µg of linear pUC19 DNA to mononucleotides in 10 minutes at 37 °C.
Storage Buffer: Baseline-ZERO DNase is supplied in a 50% glycerol solution containing 50 mM Tris-HCl (pH 7.5), 10 mM CaCl2, 10 mM MgCl2 and 0.1% Triton® X-100.
10X Baseline-ZERO™ DNase Reaction Buffer: 100 mM Tris HCl (pH 7.5), 25 mM MgCl2 and 5 mM CaCl2.
10X Baseline-ZERO™ DNase Stop Solution: 30 mM EDTA.
Quality Control: Baseline-ZERO DNase is assayed for its ability to completely digest linear dsDNA to mononucleotides under standard assay conditions. Baseline-ZERO DNase is free of detectable RNase activities as assayed by PAGE analysis of 1 µg of a synthetic RNA transcript following an overnight incubation with sufficient DNase I to completely digest 1,000 µg of DNA.
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