TriQuick Total RNA Extraction Reagent (TriQuick Reagent) Operating Instructions

TriQuick Total RNA Extraction Reagent (TriQuick Reagent)
Item No.: R1100
Specifications: 100ml
Storage: Store at 4°C in the dark and have a shelf life of 12 months.
Product introduction:
This reagent is a universal total RNA extraction reagent suitable for the extraction of total RNA from animal and plant cells or tissues and bacteria, which can effectively prevent the degradation of RNA during extraction. The obtained RNA can be directly used for a series of operations such as Northern hybridization, purification of mRNA, in vitro translation, RNase protection assay, 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 freshly opened or special chloroform, isopropanol, 75% ethanol, DEPC treated water.
1. Cell lysis or tissue homogenization: 1) Adherent cells: Exhaust the culture medium, add 1 ml TriQuick Reagent to cells per 10 cm 2 (6-well plate or 35 mm plate), cover the culture cells, and then use a straw. Or pipette 2 to 3 times, the cells should be completely lysed and then transferred to a centrifuge tube. 2) Suspended cells: Collect cells by centrifugation, drain the liquid, add 5ml to 10 million (5 × 10 6 -1 × 107) animals or yeast cells, or 10 million (1 × 107) bacteria, add 1ml TriQuick Reagent. Blow it with a pipette or pipette to completely lyse it. Some yeasts and bacteria, such as insufficient lysis, can be homogenized with a homogenizer to ensure complete lysis. Transfer to the centrifuge tube. 3) Tissue: The tissue is first cut into small pieces and placed in a glass homogenizer. The frozen tissue can be ground in a mortar. 1 ml of TriQuick Reagent was added per 50-100 mg of tissue and homogenized to complete lysis. Transfer to the 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 matter remains after homogenization, and can be centrifuged at 12,000 g for 10 minutes at 4 ° C, and then the supernatant is aspirated into a new centrifuge tube.

2. Separation: Add 0.2 volume of chloroform (1 ml TriQuick Reagent to 0.2 ml of chloroform) to the centrifuge tube containing the lysate, mix well by shaking for 30 seconds on the shaker, and let stand for 2-3 minutes at room temperature. Centrifuge at 12000g for 10 minutes at 4°C, then pipette the upper aqueous phase containing total RNA into a new centrifuge tube and draw approximately 0.5-0.6 ml per ml of TriQuick Reagent. The organic and intermediate layers contain DNA and protein and should be avoided.

3. Precipitation: Add 0.5 ml of isopropanol per ml of TriQuick Reagent, mix by inversion several times, and precipitate at room temperature for 10 minutes. After centrifugation at 12000 g for 10 minutes at 4 ° C, RNA precipitation was observed at the bottom of the tube. Discard the supernatant, add 1 ml of 75% ethanol per ml of TriQuick Reagent, and gently mix by inversion 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. Allow to dry for 5 to 10 minutes at room temperature. 4. Dissolution: Add appropriate amount of DEPC to treat the water to dissolve the RNA precipitate. Store at -80 °C.

Note :
1. The vessels, centrifuge tubes, and tips used in the RNA extraction process should be free of contamination and no RNase. Care should be taken during the operation to prevent exogenous RNase from contaminating the sample and causing degradation of the RNA sample. 2, the reagent contains harmful substances such as phenol, pay attention to personal protection.
Related Products:
R1600 DEPC treated water
P1011 Phenol: Chloroform: Isoamyl alcohol = 25:24:1 (PH<5.0)
R1050 5×RNA Loading Buffer
M1010 10×MOPS buffer
SR0020 RNAwait (non-frozen tissue RNA preservation solution)

API (Active Pharmaceutical Ingredients)

API is prepared by chemical synthesis, plant extraction or biotechnology, but which can not be taken directly by patients, are generally made into drugs that can be used directly by adding excipients and processing. Currently, China and India are the two major producers of API.