Description
DNase set for on-column DNA removal
RNase-Free DNase Set is an RNase-free DNase I set for on-column or in-solution removal of genomic DNA during RNA purification. It is supplied with reaction buffer for efficient, reproducible elimination of DNA contamination ahead of RT-qPCR and other RNA applications.
The set contains DNase I and Buffer RDD. For on-column treatment, mix 10 μl of DNase I with 70 μl of Buffer RDD, apply the 80 μl working solution to the spin column after the first wash, and incubate for 15–25 min at room temperature before continuing with your RNA purification protocol. It fits into common spin-column RNA extraction workflows for cells, animal tissues and plant tissues, giving DNA-free RNA with minimal RNA degradation.
Each set provides 50 reactions. For research use only.
Key Features
| Efficient gDNA removal | Removes genomic DNA during RNA extraction. | |
| DNA-free RNA | Ensures DNA-free RNA for accurate downstream analysis such as RT-qPCR. | |
| High specificity | Removes DNA with minimal RNA degradation. | |
| Workflow compatible | Fits into common spin-column RNA extraction workflows. |
Specifications
| Enzyme | DNase I, RNase-free |
| Application | Removal of genomic DNA during RNA purification |
| Working solution | 10 μl DNase I + 70 μl Buffer RDD per reaction |
| Incubation | 15–25 min at room temperature (20–30°C) |
| Shelf life | 2 years |
Components
| Components | 50 rxns |
| DNase I | 550 μl (1550 U) |
| Buffer RDD | 4 ml |
Storage
Store at -30 ~ -15°C.
On-column DNase treatment, performed within the steps of a spin-column RNA purification kit.
- Prepare the DNase I working solution
- For each reaction, add 10 μl of DNase I to 70 μl of Buffer RDD and mix gently. Prepare fresh.
- First wash
- Following your RNA extraction kit protocol, add 350 μl of wash buffer to the RNA spin column with bound RNA.
- Centrifuge at 12,000 rpm (13,400 × g) for 30 sec and discard the filtrate.
- DNase digestion
- Add 80 μl of DNase I working solution to the centre of the column membrane.
- Incubate at room temperature (20–30°C) for 15–25 min.
- Second wash
- Add 350 μl of wash buffer to the column, centrifuge at 12,000 rpm (13,400 × g) for 30 sec and discard the filtrate.
- Continue with the remaining steps of your RNA extraction kit (further washes and elution).
Q1: How do I prepare the DNase I working solution?
- For each reaction, add 10 μl of DNase I to 70 μl of Buffer RDD and mix gently. Prepare it fresh before use.
Q2: At what point in RNA purification is the DNase step performed?
- After the RNA is bound to the spin column and washed once with 350 μl of wash buffer. Apply 80 μl of working solution to the membrane, incubate 15–25 min at room temperature (20–30°C), wash again with 350 μl of wash buffer, then continue with the kit protocol.
Q3: What is in the set?
- The 50-reaction set contains 550 μl of DNase I (1550 U) and 4 ml of Buffer RDD.
