Navigation Links
The Fastest, Simplest, and Most Effective Way to Remove DNA Contamination

TURBO DNA-free Kit
Orders of magnitude better DNA removal than with wild type DNase I

Perform DNA digestion and sample clean-up in just 20 minutes

Preserve RNA quality

DNase treatment is the method of choice for the removal of contaminating genomic DNA from RNA samples in preparation for RT-PCR. TURBO DNase (patent pending) is an engineered variant of DNase I with a 6-fold lower Km for DNA than wild type DNase I. As a result, TURBO DNase is a superior enzyme for removal of trace amounts of contaminating DNA from RNA preparations. Using the TURBO DNA-free Kit, DNA digestion and the subsequent removal of DNase and divalent ions from the RNA sample, can be accomplished within 20 minutes.

We compared the TURBO DNA-free Kit with a popular wild type DNase I enzyme (Competitor P). To highlight the benefits of TURBO DNase, total RNA was purified from mouse spleen, a notoriously DNA-rich tissue, using a procedure that results in significant contamination with genomic DNA. An 8 g sample of spleen RNA was then digested for 15 min at 37C with 12 U of TURBO DNase or 12 U of DNase I from Competitor P in a 100 l reaction. Reactions were then processed as instructed by the TURBO DNA-free Kit Instruction Manual o r by the competitors protocol. The treated RNA was subjected to real-time PCR to detect genomic DNA contamination that remained after DNase treatments. A GAPDH primer/probe set was used in triplicate reactions, and the extent of genomic DNA removal was calculated from the real-time PCR cycle threshold (Ct) values.

Elimination of DNA contamination by TURBO DNase, compared to no DNase treatment, shifted the real-time PCR results by >20 Ct values, representing >1.2 million fold reduction of DNA contamination. When compared to standard DNase I treatment, TURBO DNase decreased genomic DNA by an additional 22.6 fold (Figure 1). Indeed, one of the three TURBO DNA-free replicates was negative even after 40 cycles of PCR.

Figure 1. The TURBO DNA-free Kit Removes >20-fold more Genomic DNA than Competing Wild Type DNase I. RNA was isolated from mouse spleen using a procedure that results in significant contamination with genomic DNA. 8 g of the RNA was then treated with either 12 U of TURBO DNase or DNase I from Competitor P in a 100 l reaction. After DNase digestion, samples were processed by following the TURBO DNA-free or the competitors protocol. Triplicate samples of the treated RNA were subjected to real-time PCR using a GAPDH primer/probe set. Note that treatment with TURBO DNase shifted the average Ct threshold 4.5 cycles, equivalent to 22.6 fold reduction of genomic DNA contamination.


By comparison, the wild type DNase I removed 20 times less DNA; GAPDH was detected in all three replicates after 30 cycles. In addition, RNA LabChip analysis of the DNase-treated RNA revealed that TURBO DNA-free was far superior in preserving RNA quality (28S/18S rRNA=1.1) than the competitors method (28S/18S rRNA=0.36). The TURBO DNA-free Kit safeguards RNA quality using reagents that are more potent and much faster and simpler to use than competing products that include wild type DNase I.


Scientific Contributors

Jon Kemppainen, Gary Latham Ambion, Inc.


back to top


Ordering Information
Cat# Product Name Size 1907 TURBO DNA-free 50 rxns
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. DNA-free A NEW Method to Remove DNA
2. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
3. Preventing Carry-over Contamination with Uracil-DNA Glycosylase
4. Combine High Yield, Great Accuracy and the Prevention of Carry-over Contamination by Using the Novel Expand High FidelityPLUS PCR System
5. Combat RNase Contamination in the Lab
6. Testing Compressed Air Lines for Microbiological Contamination
7. Testing Compressed Air Lines for Microbiological Contamination
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:9/12/2017)... ... , ... It wasn’t just the San Diego temps that soared this summer. ... has had a record-breaking number of guests for corporate cooking challenges, team events and ... company outings, family get-togethers, anniversaries and milestone birthdays. While all groups gathered in ...
(Date:9/12/2017)... ... 2017 , ... September 15, 2017. Pittsburgh, PA. The ... the Department of Surgery and Cancer and Director of the MRC-NIHR National Phenome ... His presentation, “Analytical Science in Precision Medicine: Facing the Challenges of the 21st ...
(Date:9/12/2017)... ... 2017 , ... Facet Life Sciences, a leading provider of development solutions (software ... PhD has joined the company’s Board of Directors. , Dr. Massa brings 38 ... Most recently he served as VP and Head of Global Regulatory Operations at Sanofi. ...
(Date:9/12/2017)... ... September 11, 2017 , ... As part of BD’s ... BD is sponsoring an educational webinar in recognition of World Sepsis Day ... estimated five million deaths annually, is preventable and treatable through increased awareness and ...
Breaking Biology Technology:
(Date:4/5/2017)... LONDON , April 4, 2017 KEY ... is anticipated to expand at a CAGR of 25.76% ... neurodegenerative diseases is the primary factor for the growth ... full report: https://www.reportbuyer.com/product/4807905/ MARKET INSIGHTS The ... of product, technology, application, and geography. The stem cell ...
(Date:3/30/2017)... YORK , March 30, 2017 Trends, ... type (physiological and behavioral), by technology (fingerprint, AFIS, iris ... voice recognition, and others), by end use industry (government ... and immigration, financial and banking, and others), and by ... Europe , Asia Pacific , ...
(Date:3/24/2017)... Mar 24, 2017 Research and Markets has ... Market Analysis & Trends - Industry Forecast to 2025" report ... ... at a CAGR of around 15.1% over the next decade to ... report analyzes the market estimates and forecasts for all the given ...
Breaking Biology News(10 mins):