RNA Extraction Reagent
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Price:
US$362.50
(Size: 100 ml)
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Abbexa's RNA Extraction Reagent is a rapid, quick and sensitive method for extracting total RNA from a variety of cells and tissues. The reagent combines phenol and guanidine thiocyanate in a mono-phase solution to inhibit RNase. After lysis and centrifugation, the solution seperates into three phases: an aqueous phase containing RNA and an interphase and a pink organic phase. RNA can be precipitated by addition of isopropanol.
This product also comes with 15 ml of RNA Dissolving Solution.
| Target |
RNA Extraction Reagent |
| Storage |
Store in the dark at 2-8 °C. Stable for 12 months from date of receipt. |
| Availability |
Shipped within 10-20 working days. |
| Note |
This product is for research use only. |
| Directions for use |
Sample Preparation - Adherent cells: Wash the culture dish with 1X PBS and detach cells using a cell spatula. Add 1 ml of RNA Extraction Reagent per 10 cm3 culture dish and lyse cells by pipetting up and down. Collect the lysate in a microcentrifuge tube and allow to stand at room temperature for 5 min.
- Suspension cells: Collect the suspension cells in a microcentrifuge tube and centrifuge at 8000 × g at 2-8 °C for 2 min. Discard the supernatant and add 1 ml of RNA Extraction Reagent per 107 cells. Pipette up and down to remove any precipitates from lysate and allow to stand at room temperature for 5 min.
- Tissue homogenates: Weigh the tissue sample and grind completely to make a tissue powder using liquid nitrogen. Collect the completely grinded tissue powder in a microcentrifuge tube and for each 50-100 mg tissue, add 1 ml of RNA Extraction Reagent. Homogenize using a homogenizer and repeatedly pipette up and down. Allow the tissue homogenate to stand at room temperature for 5 min.
Assay Procedure - Add 0.2 ml of Chloroform per ml of RNA Extraction Reagent used. Shake the tube vigorously for 30 sec and allow to stand at room temperature for 3 min.
- Centrifuge the tube at 10000 × g at 2-8 °C for 15 min. The solution thus obtained seperates into three different layers: lower pink organic phase, an interphase and a colorless upper aqueous phase which contains the RNA.
- Collect the colorless upper phase layer containing the RNA in a RNase-free tube.
- Add 0.5 ml Isopropanol for per ml of RNA Extraction Reagent used. Mix thoroughly by inverting the tube and allow to stand a room temperature for 10 min.
- Centrifuge at 10000 × g at 2-8 °C for 10 min. Discard the supernatant.
- Add 1 ml of 75% ethanol (prepared with RNase-free water) per 1 ml of RNA Extraction Reagent used. Vortex vigorously.
- Centrifuge at 7500 × g at 2-8 °C for 5 min. Discard the supernatant and air dry the RNA pellet for 5 min.
- Dissolve the RNA pellet in 50-100 µl of RNA Dissolving Solution and incubate at 55-60 °C for 10 min or at -70 °C for long term storage.
Note - Ensure the complete extraction by sufficiently shaking the tube after adding Chloroform.
- All other reagents and consumbles like Isopropanol, 75% Ethanol, which are not provided, should be RNase-free.
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