RNA Extraction Solution

RNA Extraction Solution
For the extraction of total RNA from cells or tissues, RNA with high integrity and purity can be obtained

Product Information

Product Introduction
RNA Extraction Solution is a ready-to-use reagent, designed to isolate total RNA from cell and tissue samples. It utilizes substances such as guanidine isothiocyanate to rapidly lyse the cells, causing RNA to be released and extracted into phenol, and it also contains an RNase inhibitor, which prevents the degradation of RNA and maintains the integrity of RNA.

Storage and Shipping Conditions
Ship with ice packs; Store in the dark at 2-8; valid for 12 months.

Preparation for the experiment
Materials required but not provided：
Alternative to chloroform (G3014), isopropanol, DEPC water or RNase-free water (G4700), 75% ethanol (prepared with DEPC-treated water). 

Assay Protocol / Procedures
1. Lyse samples Adherent cells: Discard the cell-culture medium, wash the cells with pre-cooled PBS. Add 1 mL of RNA Extraction Solution to each well of a 6-well plate and 0.5 mL of RNA extraction solution to each well of a 12-well plate. Pipet the lysate up and down several times to homogenize, transfer them to a new tube and incubate for 5 minutes at room temperature .
Suspension cells: Collect the cells by centrifugation, and discard the supernatant completely. Add 1 mL of RNA Extraction Solution per 1-10×106 cells to lyse the cells, pipet the lysate up and down several times to homogenize, and incubate for 5 minutes at room temperature 
Tissue sample: Add 1 mL of RNA Extraction Solution per 100 mg of tissue to the sample, and homogenize until there are no visible tissue clumps. Centrifuge at 12,000 × g for 10 min at 4°C and transfer the clear supernatant to a new tube.
Blood samples：Fresh blood samples were added with 800 μL of RNA extraction solution per 200 μL, pipette gun was blown repeatedly to mix, and left to stand at room temperature for 5-10 min; in the case of blood samples preserved in Tissue RNA Stabilization and Preservation Solution (G3019), the samples were first centrifuged at 4000-5000 rpm for 5 min, and the upper layer of Tissue RNA Stabilization and Preservation Solution was discarded, and then 800 μL of RNA Extraction Solution was added, and the pipette gun was blown repeatedly to mix the samples, and then the samples were allowed to stand at room temperature for 5-10 min.
2. RNA Extraction: Add 100 μL of chloroform substitute (G3014) per 1 mL of RNA extract, vortex mix (recommended vortex mixer MV-4500B) or centrifuge tube up and down with vigorous shaking for 15 s, and let it stand at room temperature for 3-5 min.
3. Centrifugation：Centrifuge at 12,000 × g for 15 minutes at 4°C, Carefully transfer 500-550 µL colorless upper aqueous phase to a new tube, per 1 mL of RNA Extraction Solution used for lysis.
4. Precipitate the RNA: Add 550 µL of isopropanol to the aqueous phase, invert and gently rotate the tube several times, and incubate at room temperature or 4°C for 10 min (or -20°C for 15 min).
5. Centrifugation：Centrifuge at 12,000 × g for 10 minutes at 4°C. Total RNA precipitate forms a white gel-like pellet at the bottom of the tube. Discard the supernatant.
6. Wash: Add 1 mL of 75% ethanol and mix upside down (until a white precipitate floats preferably).Centrifuge at 12,000 × g for 5 min at 4°C, discard the supernatant, briefly centrifuge at high speed (5,000 × g for 3-5 seconds), and carefully remove any remaining solution.
7. Solubilize the RNA: Leave the centrifuge tube with RNA precipitation open for 3-5 min to allow the RNA to dry slightly, then add 20-50 μL of DEPC water or RNase-free water (or use RNA solubilizing solution G3029, G3030) to fully solubilize the RNA, and then it can be used for concentration assay and subsequent experiments, and the RNA needs to be preserved at -70  if it is impossible to carry out the subsequent experiments in a timely manner. Be careful not to over-dry the RNA or it will be difficult to dissolve and will affect the A260/280 ratio.

Notes:
1. All pipette tips and centrifuge tubes need to be free of RNAase contamination and can be purchased as our sterile, enzyme-free consumables.
2. This reagent contains guanidine isothiocyanate and redistilled phenol, avoid contact with skin or inhalation.
1. For your safty and health,please wear safety glasses, gloves, or protective clothing.