RTS (Room Temperature Stable) DNase is a highly purified DNase I enzyme formulated in a unique stabilization solution that provides long term stability at room temperature. The RTS DNase™ Kit is used for the removal of genomic DNA contamination in RNA preparations. RTS DNase will remove up to 30 µg of DNA in 20 minutes using 10 units (1 µl) of enzyme. The enzyme is stable for up to 6 months at room temperature with no loss of activity and for 2 years at 4°C without loss of activity. The RTS DNase™ Kit also contains a novel and highly specific resin which is used to bind and remove the RTS DNase enzyme and divalent cations from the reaction, eliminating the need for heat or EDTA inactivation of the DNase. The RNA is ready to use immediately after resin treatment.
Preserve enzyme activity
RTS DNase is the first DNase I enzyme which is stable at room temperature, so there is no need to aliquot and freeze stocks of the enzyme. Room temperature stability eliminates concern about freeze-thaw cycles that may decrease enzymatic activity. RTS DNase maintains the highest activity over the life of the kit, enabling consistent results with all RNA samples.
Protect your RNA
Isolating high quality RNA is a time-consuming and expensive process. Consequently, it is essential to prevent RNA degradation during DNase treatment. The RTS DNase™ kit protects valuable RNA samples during this process, as it contains only Certified RNase-free reagents, and does not require heat, EDTA or harsh chemicals.
The RTS DNase™ Kit protocol starts with dilution of RTS DNase and 10X RTS DNase buffer to a final concentration of 1X in the digestion reaction. The reaction is then incubated at 37°C for 20 minutes. Removal of the DNase and divalent cations is performed by adding RTS DNase Removal Resin and incubating for 10 minutes at room temperature. The resin is pelleted by centrifugation, and the RNA sample is transferred to a new tube, ready for use in RT-PCR and further analysis.
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Fig 1. RTS DNase eliminates genomic DNA in RNA. RNA was prepared from two different soils using the RNA PowerSoil® Total RNA Isolation Kit, followed by DNase treatment with the RTS DNase™ Kit. Agarose gel analysis demonstrates the efficient removal of genomic DNA from the RNA sample.
Fig 2. Quantification of DNA levels in RNA before and after RTS DNase treatment. The RNA samples described in Fig. 1 were analyzed using 16S rRNA gene universal primers and a one-step qRT-PCR kit before and after RTS DNase treatment. To quantify the level of reduction of genomic DNA, qRT-PCR was performed using 1 µl of the treated or untreated RNA. Genomic DNA was reduced 5 logs (>15 cycles) and is below the level of background DNA in the qPCR Kit (red line). The standard curve (grey lines) demonstrates an assay efficiency of 99%.
Fig 3. RTS DNase Removal Resin completely removes DNase. Samples were subjected to DNase treatment and enzyme removal using the RTS DNase™ Kit or a competitor’s kit according to the manufacturer’s protocols, and then analyzed for residual DNase activity using the MO BIO DNase-free certification assay. Lane 5 is the negative control and did not receive DNase. Samples were incubated for 1 hour at 37oC, followed by inactivation for 5 minutes at 65oC. Results are shown on a 1% agarose gel. The RTS DNase Removal Resin successfully removed the DNase (lanes 3-4), while the competitor’s resin failed to remove all of the DNase from the samples (lanes 1-2).
|Unit Definition||10 units/µl, removes 30µg DNA|
|Stability||6 months at room temperature|
|2 years at 4ºC|
|Equipment Recommended||Vortex Genie® 2 Vortex|
|Vortex adapter for 2 ml tubes|
Store the RTS DNase™ Kit at 4ºC for up to 2 years, or at room tempearture for up to 6 months.
|RTS DNase||55 µl|
|RTS DNase Removal Resin||550 µl|
|10X RTS DNase Buffer||550 µl|
|RNase-free Water||2 x 1 ml|