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| Product Name | DNA-Free DNA Removal Kit |
| Catalog No. | NATR-HMM-0134 |
| Description | A post-RNA-purification kit for the complete removal of contaminating genomic DNA from RNA preparations without compromising RNA integrity. The kit uses a recombinant DNase I treatment followed by a novel DNase removal resin that eliminates the enzyme without heat inactivation, organic extraction, or precipitation. |
| Intended Use | Removal of trace genomic DNA contamination from RNA samples prepared for highly sensitive applications including RT-qPCR with intron-spanning primers, RNA-seq library construction, microarray analysis, and any application where even minimal DNA carryover would produce false-positive results. |
| Principle / Technology | Recombinant RNase-free DNase I digests both single-stranded and double-stranded DNA in the RNA sample. A proprietary DNase inactivation resin then selectively binds and removes the DNase I enzyme, divalent cations, and digested DNA fragments from the solution. The resin is pelleted by centrifugation, leaving RNA in the supernatant without the need for heating, phenol extraction, or alcohol precipitation steps. |
| Detection Method | qPCR with human genomic DNA-specific primers (e.g., repetitive elements, single-copy intronic sequences) to verify DNA removal; agarose gel electrophoresis to confirm RNA integrity; RT-minus controls in RT-qPCR as internal validation for each sample; Bioanalyzer RNA integrity assessment before and after treatment |
| Sample Type | Total RNA purified by any method (silica column, phenol extraction, lithium chloride precipitation); RNA from mammalian, plant, bacterial, yeast, and viral sources; partially degraded RNA from FFPE and archival samples |
| Performance Range / Specifications | Removes >99.9% of genomic DNA from up to 10 μg total RNA per treatment; RNA recovery: >90% of input RNA; treatment time: <30 minutes; RNA fragment size: all RNA species preserved including small RNAs (<200 nt) and mRNAs up to >10 kb |
| Sensitivity / LOD | Reduces genomic DNA to undetectable levels by 40-cycle qPCR using single-copy gene primers from 1 μg treated RNA; no DNA amplification in RT-minus qPCR control at input equivalent to 100 ng cDNA synthesis |
| Specificity | Recombinant DNase I specific for DNA substrates; no detectable RNase or non-specific ribonuclease activity after 4-hour incubation with total RNA; the inactivation resin specifically removes DNase without binding RNA |
| Reaction Conditions / Protocol | For each 50 μL RNA sample (up to 10 μg RNA), add 5 μL 10× DNase I buffer and 1 μL rDNase I (2 U), incubate at 37°C for 20–30 minutes, add 5 μL (or 0.1 volume) DNase inactivation resin, incubate at room temperature for 2 minutes with periodic mixing, centrifuge at 10,000g for 1.5 minutes, carefully transfer supernatant containing DNA-free RNA to a fresh tube without disturbing the resin pellet; total time approximately 25 minutes |
| Components / Formulation | Recombinant DNase I (rDNase I, 2 U/μL), 10× DNase I buffer (100 mM Tris-HCl pH 7.5, 25 mM MgCl2, 5 mM CaCl2), DNase inactivation resin (proprietary slurry formulation), nuclease-free water |
| Storage Conditions | DNase I and buffer at –20°C; inactivation resin at –20°C or 2–8°C (do not freeze resin beyond –20°C); rDNase I sensitive to physical denaturation — mix gently by flicking, do not vortex; aliquot enzyme to avoid repeated freeze-thaw |
| Shelf Life | 18 months at –20°C from date of manufacture; inactivation resin stable for 12 months at 2–8°C after opening |
| Package Specifications | 50 reactions (based on 50 μL per reaction, up to 10 μg RNA each), 200 reactions |
| Product Form | Liquid enzyme and buffer; slurry resin suspension |
| Quality Control | Each lot tested: genomic DNA removal from 10 μg HeLa total RNA — no Cq in 40-cycle qPCR for Alu repeats and GAPDH intronic primer sets; RNA recovery >90% measured by fluorometric quantitation; RIN score unchanged before and after treatment; DNase activity verified by plasmid digestion assay; no residual DNase activity in treated RNA after resin treatment |
| Key Features | No heat inactivation required — preserves RNA integrity for RNA-seq; resin-based DNase removal eliminates phenol extraction and ethanol precipitation; superior to EDTA heat-inactivation methods for RT-qPCR sensitivity; rapid 25-minute protocol; demonstrated performance with FFPE and low-quality RNA samples |
For research use only, not for clinical use.
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