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G3013-100ML RNA Extraction Liquid Molecular Biology Reagent

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G3013-100ML RNA Extraction Liquid Molecular Biology Reagent

RNA extraction solution is a ready-to-use reagent, designed to isolate high quality total RNA from cell and tissue samples.

RNA extraction solution is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components which facilitate the isolation of a variety of RNA species of large or small molecular size.

RNA extraction solution maintains the integrity of the RNA due to highly effective inhibition of RNase activity while disrupting cells and dissolving cell components during sample homogenization.

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Technical Specification

Storage and Handling Conditions

Wet ice bag transportation; Stored at 4ºC in the dark; valid for 12 months.
 

Preparation for the experiment

Bring your own solution as follows:

Chloroform, isopropanol, DEPC water, 75% ethanol (prepared with DEPC water)
 

Assay Protocol / Procedures

Lysis of cell or tissue sample

Adherent cells: Remove the cell culture medium, wash the cells with an appropriate amount of pre-cooled PBS, and try to remove the PBS as much as possible. 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. Blow the cells with a gun to lyse the cells, then transfer them to a centrifuge tube and let stand at room temperature for 5 min.

Suspension cells: collect cells by centrifugation, and remove the supernatant as much as possible. Add 1 mL of RNA extract for every 1-10×106 cells, pipette several times to lyse the cells, and let stand at room temperature for 5 minutes.

Tissue sample: Cut the tissue into small pieces, put it in a pre-cooled homogenizer, add 1 mL of RNA extraction solution per 100 mg of tissue, and homogenize until there are no visible tissue clumps. Centrifuge at 12000 g for 10 min at 4°C to separate the tissue fragments, and transfer the supernatant to a new centrifuge tube.

  1. RNA extraction: add 380µL of chloroform to each 1 mL of RNA extraction solution, vortex to mix or shake the centrifuge tube upside down vigorously for 15 s, and let it stand at room temperature for 3 min.
  2. Centrifugation: Centrifuge at 12000 g for 15 min at 4°C. Carefully transfer the upper colorless water phase to a new centrifuge tube. Approximately 500-550µL per ml of extract can be drawn.
  3. Precipitate RNA: Add 550µL of isopropanol to the tube of aqueous extract collected above, gently invert and mix several times, and precipitate at -20°C for 15 minutes.
  4. Centrifugation: Centrifuge at 12000 g for 10 min at 4°C. A white precipitate at the bottom of the tube is total RNA. Discard the supernatant.
  5. Washing: Add 1 mL of 75% ethanol and mix by inversion. Centrifuge at 12000 g for 5 min at 4°C, discard the supernatant, and then briefly centrifuge at high speed (5000 g for 3-5 s), and carefully suck up the liquid with a micropipette.
  6. RNA dissolution: Leave the opening of the centrifuge tube with RNA precipitation for 3-5 min. After the RNA is slightly dried, add 20µL of DEPC water to fully dissolve the RNA, which can be used for concentration detection and follow-up experiments. The RNA must be stored at -70°C during the experiment. Be careful not to let the RNA dry too much, otherwise 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 RNase contamination. Sterile non-enzyme consumables can be purchased from our company.
  2. Avoid skin contact or inhalation because this reagent contains guanidine isothiocyanate and redistilled phenol.
  3. Please wear lab coat and disposable gloves during operation.
     

For research only, not for clinical diagnosis!

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