Quantity :100ml

Storage: 2-8℃. Keep in Dark Place

 

Shelflife: 12 months

Transport temperature:Transport at normal temperature

Product introduction:
This reagent is a universal total RNA extraction reagent, suitable for Total RNA extraction from animal and plant cells or tissues and bacteria can effectively prevent RNA degradation during the extraction process. The obtained RNA can be directly used for a series of operations such as Northern hybridization, purified mRNA, in vitro translation, RNase protection experiments, RT-PCR and cDNA cloning. This reagent can be used for 100 total RNA extractions (10 square centimeters of cells or 100 mg of tissue).
Operating Instructions:
Bring your own newly opened or special chloroform, isopropyl alcohol, 75% ethanol, and DEPC treated water.
1. Cell lysis or tissue homogenization:
1) Adherent cells: Aspirate the culture medium and add 1 ml TriQuick Reagent per 10 square centimeters (6-well plate well or 35 mm dish) of cells. Cover the cultured cells and pipet 2 to 3 times with a pipette or pipette. The cells should be completely lysed and then transferred to a centrifuge tube.
2) Suspension cells: Collect cells by centrifugation and absorb all the liquid. Every five million to ten million (5×106-1×107) animal, plant or yeast cells, or ten million (1×107) bacteria , add 1ml TriQuick Reagent. Use a pipette or pipette to pipet until complete lysis. If certain yeasts and bacteria are not fully lysed, they can be homogenized with a homogenizer to ensure complete lysis. Transfer to centrifuge tube.
3) Tissue: First cut the tissue into small pieces and put them into a glass homogenizer. Frozen tissue can be homogenized in a mortar. Add 1ml TriQuick Reagent per 50-100mg of tissue and homogenize until completely lysed. Transfer to centrifuge tube.
The cleavage product should be a clear, transparent viscous liquid. Leave at room temperature for 5 minutes. For tissue samples rich in impurities such as polysaccharides and proteins, insoluble substances will still remain after homogenization. You can centrifuge at 12,000g and 4°C for 10 minutes, and then transfer the supernatant to a new centrifuge tube.
2. Separation: Add 0.2 times the volume of chloroform to the centrifuge tube containing the lysate (add 0.2 ml chloroform to 1 ml TriQuick Reagent), shake and mix thoroughly on a shaker for 30 seconds, and leave it at room temperature for 2-3 minutes. . Centrifuge at 12000g for 10 minutes at 4°C, then pipet the upper aqueous phase containing total RNA into a new centrifuge tube. Approximately 0.5-0.6 of each ml of TriQuick Reagent can be pipettedml. The organic phase and middle layer contain DNA and proteins and should be kept out of reach.
3. Precipitation: Add 0.5 ml of isopropyl alcohol per ml of TriQuick Reagent, mix by inverting several times, and precipitate at room temperature for 10 minutes. Centrifuge at 12000g for 10 minutes at 4°C. RNA precipitate will be visible at the bottom of the tube. Discard the supernatant, add 1 ml of 75% ethanol per ml of TriQuick Reagent, and mix gently by inverting to wash the RNA pellet. Centrifuge at 12000g for 2 minutes at 4°C, discard the liquid, and be careful not to discard the RNA pellet. Place upside down to dry at room temperature for 5 to 10 minutes.
4. Dissolution: Add an appropriate amount of DEPC treated water to dissolve the RNA precipitate. Store at -80℃.
Notes:
1. The utensils, centrifuge tubes, pipette tips, etc. used in the RNA extraction process should be free of contamination and RNase. Care should be taken during operation to prevent exogenous RNase contamination of the sample and degradation of the RNA sample. 2. The reagent contains harmful substances such as phenol, so please pay attention to personal protection.

 

Remarks:

The above data are all from public literature and have not been independently verified by Sunlong Biotech. They are for reference only.

These protocols are for reference only. Sunlong Biotech does not independently validate these methods.

 

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