Ambion® TURBO DNA-free™ contains reagents for the efficient, complete digestion of DNA along with the removal of the enzyme and divalent cations post-digestion. The kit includes sufficient reagents for 50 rxns.
Features of Ambion® TURBO DNA-free™ include:
• Hyperactive TURBO™ DNase is a catalytically superior enzyme compared to wild type DNase I
• Removes trace quantities of DNA that can plague RT-PCR
• Reagent included to completely remove DNase without phenol treatment or heating
The Best Method for Genomic DNA Removal Prior to RT-PCR
TURBO™ DNase is a recombinant, engineered form of DNase I that is much more efficient than wild type DNase I in digesting away trace amounts of unwanted DNA. TURBO™ DNase binds DNA substrates 6-fold more tightly than traditional DNase I, making this enzyme the tool of choice for clearing residual DNA that can generate a false positive signal in RT-PCR applications. TURBO™ DNase now includes an enhancer that increases the effectiveness by two orders of magnitude.
Efficient DNase and Divalent Cation Removal Without Organic Extraction or Precipitation
Conventional DNase treatment procedures of RNA samples prior to RT-PCR typically call for inactivation of the DNase by phenol:CHCl3 extraction or heating followed by a precipitation step to concentrate the RNA. Phenol:CHCl3 extractions can be cumbersome and time-consuming. Heating the sample to inactivate DNase can lead to chemical degradation of the RNA by divalent cations present in the DNase buffer. The TURBO DNA-free™ Kit circumvents these problems using a novel DNase Inactivation Reagent. In addition to removing the TURBO™ DNase from the reaction, the Inactivation Reagent also binds and removes divalent cations from the TURBO™ DNase Reaction Buffer.
Accessory Product:
TURBO™ DNase alone is available in 1,000 U (AM2238) and 5,000 U (AM2239) pack sizes.
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![Complete removal ...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/f00761.jpg","Complete removal of divalent cations with the TURBO DNA-free™ Inactivation Reagent.","100 ng of total HeLa-S3 RNA was TURBO DNase treated followed by DNase removal with the DNase Inactivation Reagent from the TURBO DNA-free™ Kit. The samples were either heated at 75°C for 10 min or at 95°C for 5 min. Untreated samples were resuspended in water and heated at 75°C for 10 min or not heated. In addition, a 100 ng untreated RNA sample was resuspended in 1X DNase Reaction Buffer and heated at 75°C for 10 min (lane 5). All samples were loaded on an RNA 6000 LabChip® (Caliper Technologies) and run on an Agilent bioanalyzer 2100. The results are presented in the form of a gel. Extensive degradation of the non-resin–treated, heated RNA sample (lane 5) is due to chemical hydrolysis of the RNA catalysed by the divalent cations (Mg<sup>++</sup>) present in DNase reaction buffer.");)
![TURBO DNA-free&am...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/f01043.jpg","TURBO DNA-free™ reagent improves DNA removal by more than 600-fold.","Mouse spleen total RNA samples, highly contaminated with DNA (30% RNA and 70% DNA; 23 µg), were treated with 4 U of TURBO DNase in a 60 µL reaction for 20 min at 37°C, or were left untreated. DNase digestion was halted by adding 6 µL (1/10 volume) of DNase Inactivation Reagent. 2 µL of each treated sample were amplified in a 25 µL RT-PCR using a TaqMan® primer probe set for mouse GAPDH. RT-PCR analysis of the DNase-treated samples unmasked the RNA-only signal, which appeared at 15.3 C<sub>t</sub>.");)
