RapidOut DNA Removal Kit
New proprietary technology for convenient removal of gDNA from RNA sample in a simple two-step procedure. .
Detailní popis
Thermo Scientific RapidOut DNA Removal Kit rapidly and safely removes genomic DNA from total RNA and mRNA preparations. Complete digestion of DNA and safe removal of DNase I from the digestion reaction is ensured without RNA damaging steps, such as heating or organic extraction. First, the RNA sample is treated with recombinant RNase-free DNase I to levels below the limit of detection by routine PCR. DNase I is safely removed subsequently using proprietary DNase Removal Reagent (DRR).
DRR efficiently binds DNase I and the complex is collected at the bottom of the tube by centrifugation. The purified RNA is collected as a supernatant. The RNA after RapidOut procedure is free from DNA contamination and free of DNase I. It is ready to use in different applications including end-point or real-time RT-PCR, cloning, microarrays, and Northern blotting.
Highlights
- Efficient – complete ds and ssDNA digestion and proprietary technology for DNase I removal
- Rapid – single step sufficient for complete DNase I removal
- Safe – no need for toxic organic extractions or RNA-damaging heating steps
Applications
Main Applications
- RNA isolation and RNA analysis, particularly RT-qPCR and RT-PCR (customers performing expression analysis of low transcription level genes.
- Customers performing ds-cDNA synthesis from total RNA preps.
Other applications
- Elimination of DNA from RNA for microinjections and transfection.
- Elimination of DNA from RNA prior microarray analysis.
- Elimination of DNA from RNA prior Northern blot analysis.
Includes
| Component | Amount |
|---|---|
| DNase I, RNase-free (1 U/µL) | 250 µL |
| 10X Reaction Buffer with MgCl2 | 1 mL |
| DNase Removal Reagent (DRR) | 500 µL |
| Water, nuclease-free | 2 × 1.25 mL |
| Hazardous | No |
| Quality Control | Ribonuclease Assay For DRR: No contaminating RNase was detected after incubation of 10 µL of DRR with 160 ng of 2 kb RNA transcript for 4 hours at 37°C. For DNase I: No contaminating RNase was detected after incubation of 5u of DNase I with 160 ng of 2kb RNA transcript for 4 hours at 37°C. DNase I removal efficiency RNA sample after RapidOut procedure was incubated with 1 µg of supercoiled plasmid DNA for 15 min at 37°C. Less than 20% of supercoil DNA was transformed to nicked and linear forms. Functional testing Of the kit: DRR is functionally tested in RT-qPCR amplification reaction with RNA treated with RapidOut DNA Removal Kit. Of DNase I: DNase I is functionally tested in a unit activity assay: one unit of the enzyme completely degrades 1 µg of pUC19 DNA in 10 min at 37°C. |
| Storage Condition | -20 C |
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