Introduction
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from FFPE tissue and section samples (with DNase) |
| Applications | RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNase |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | FFPE tissue sample |
| Sample amount | 6mg |
| Yield | 20μg |
| Elution volume | ≥10μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Principle
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.
Advantages
- High quality – high purity total RNA can be directly used in various sensitive downstream applications
- Fast – several samples can be extracted in 60 minutes by column method
- Safe – no phenol chloroform extraction required
- Sensitive – RNA can be recovered at the level of PG
- Efficient DNA removal – unique method to effectively remove genomic DNA
Kit Contents
| Contents | R414402 | D414403 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Micro Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Buffer DPS | 60 ml | 250 ml |
| Buffer FRL | 15 ml | 60 ml |
| Buffer RLC | 15 ml | 60 ml |
| Buffer RWC* | 10 ml | 50 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| DNase I | 600 µl | 5 x 600 µl |
| DNase Booster Buffer | 1.5 ml | 6 ml |
| Protease Dissolve Buffer | 1.8 ml | 10 ml |
| Proteinase K | 24 mg | 120 mg |
| RNase Free Water | 10 ml | 20 ml |
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Experiment Data
