Description
Kit content
| Component | 50 preps | 100 preps |
| RNSol Reagent | 50 ml | 100 ml |
| RLB | 50 ml | 100 ml |
| TWB1 (concentrate) | 17 ml | 35 ml |
| TWB2 (concentrate) | 15 ml | 27.5 ml |
| Nuclease-free Water | 5 ml | 10 ml |
| Spin Column | 50 | 100 |
| Collection Tube | 50 | 100 |
Storage
Shipment condition is checked by ROJETechnologies. After arrival, RNSol Reagent should be kept dry, at 2-8 °C and all remaining reagent should be kept at room temperature (15-25 °C). When storage condition is as directed, all reagents are stable until expiration date, as indicated on the kit box.
Intended Use
The RNall Kit provides the components and procedures necessary for purifying RNA and miRNA from cultured cells, animal tissue, PBMC and bacteria cells. Notice that, RNall is intended for molecular biology applications not for diagnostic use. We recommend all users to study RNA experiments guideline, before starting their work.
Warning and Precautions
Due to chemical material usage that may be hazardous, always make sure to wear suitable lab coat, disposable gloves, and protective eyewear. Material Safety Data Sheet (MSDS) for all products and reagents are provided. They are accessible online at ROJE Technologies
Quality Control
RNall Kit is tested against predetermined experiments on a lot-to-lot basis according to ROJETechnologies ISO-certified quality management system, to ensure consistent product quality. For your information, the results of all experiments are accessible by addressing Cat and Lot number on web at ROJE Technologies
Procedure
The RNall kit is designed for isolating all forms of RNA molecules from 0.5-100 mg tissue samples, up to 1×107 animal or bacteri cells and 1-10 ml of whole blood. Fresh or frozen samples should be lysed and homogenized. For procedure in which RNsol is used as lysis buffer, Aqueous phase is separated using chloroform. After lysing samples, the rest of procedure is the same. RNA binding to the silica membrane is achieved selectively, by the addition of Ethanol to the lysate. Contaminants removed by two specific kinds of washing buffers. Pure RNA and miRNA is finally eluted in nuclease-free water. Isolated RNA is ready to use in downstream amplification. It has A 260/ A 280 ratios of 1.9–2.3 by spectrophotometer, confirming high purity.
Equipment & Reagents to be supplied by user
- Absolute Ethanol
- 1.5 ml RNase free microcentrifuge tubes
- RNase free pipets and pipet tips
- Benchtop refrigerated microcentrifuge (while RNSol is used)
- Vortex
- Thermoblock or water bath
- TissueLyser/ Mortar and pestle (for animal tissue sample)
- Syringe and needles
- Homogenizer
Applications
The isolated RNA and miRNA can be used in many downstream applications:
- RNA Sequencing
- cDNA synthesis
- Poly A+ RNA selection
- Microarrays
- RT-PCR and real-time RT-PCR
- Northern, dot blot analysis
Recommended starting material:
To reach optimized results it is better to follow as listed below. The size of the recommended starting material to use with determined RLB, RNSol, Chloroform and Ethanol volumes, are written in Table 2 and 3.
Table 2. Appropriate sample size and amount of RLB
| Sample Type | Sample Size | Amount of RLB | |
| Animal Cells | ≤5 X106 | 350 µl | |
| Animal Cells | 5 X 106 To 1 X 107 | 600 µl | |
| Animal Not Fibrous Tissues | 0.5-10 Mg | 350 µl | |
| Animal Not Fibrous Tissues | 10-30 Mg | 600 µl | |
| Animal Fibrous Tissues | 0.5-30 Mg | 300 µl | |
| Bacteria Cells | ≤5 X108 | 500 µl | |
| Bacteria Cells | 5 X 108 – 1 X 109 | 800 µl | |
| Bacteria Cells | 1 X 109 – 2 X 109 | 1000 µl | |
| Whole blood | Up to 2 x 106 cells | 350 µl | |
| Whole blood | 2 x 106 to 1 x 107 cells | 600 µl | |
Table 3. Appropriate sample size and amount of RNSol Reagent, chloroform and Ethanol amount
| Sample Size | RNSol Reagent Amount | Chloroform Amount | |
| 10 mg animal tissue, 1 × 106 animal cells | 100 µl | 100 µl | |
| 20 mg animal tissue 2 × 106 animal cells | 200 µl | 100 µl | |
| 30 mg animal tissue 3 × 106 animal cells 0.5-1 ml whole blood | 300 µl | 200 µl | |
| 40 mg animal tissue 4 × 106 animal cells | 400 µl | 200 µl | |
| 50 mg animal tissue 5 × 106 animal cells 1-3 ml whole Blood | 500 µl | 300 µl | |
| 60 mg animal tissue 6 × 106 animal cells | 600 µl | 300 µl | |
| 70 mg animal tissue 7 × 106 animal cells 3-5 ml whole Blood | 700 µl | 400 µl | |
| 80 mg animal tissue 8 × 106 animal cells | 800 µl | 400 µl | |
| 90 mg animal tissue 9 × 106 animal cells | 900 µl | 500 µl | |
| 100 mg animal tissue 107 animal cells 5-10 ml whole Blood | 1ml | 500 µl | |
Technical Assistance
ROJETechnologies guarantees your complete satisfaction. ROJE technical support team composed of highly trained experienced scientists, who are able to troubleshoot most problems you face. Our technical support team can offer expert advice which may help you select suitable product.
Contact our technical support at any time by selecting one of these ways:
- Through our telephone and fax number available at ROJETechnologies website.
- You can submit your question directly to ROJE Technical Support Team from our website (rojetechnologies.com)
Or send your questions to this email address, Technicalsupport@Rojetechnologies.com
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