Trizoe Reagent

Product Number: RNK0101

Shipping and Storage

Trizoe can be stably stored at room temperature for 12 months. Nevertheless, for optimal results, we recommend storing in an environment of 2-8°C.  

Important Note

Toxic substances that come into contact with the skin or are accidentally swallowed can cause burns. Once in contact with the skin, wash immediately with a large amount of detergent and clear water. If you feel unwell, see a doctor and seek the correct treatment plan for phenol and other components.

Description

Trizoe reagent is a reagent that directly extracts total RNA from cells or tissues. It can maintain the integrity of RNA during cell fragmentation and lysis. After adding chloroform and centrifuging, the sample is divided into a water sample layer and an organic layer.RNA exists in the water sample layer. After collecting the water sample layer above, RNA can be reduced through isopropanol precipitation. After removing the water sample layer, the DNA and proteins in the sample can also be successively reduced by precipitation. Ethanol precipitation can precipitate DNA from the intermediate layer, while adding isopropanol to the organic layer can precipitate protein. Co purified DNA is very useful for standardizing RNA production between samples.

Whether it is human, animal, plant, or bacterial tissue, this method applies to small amounts of tissue (50-100mg) and cells (5×10⁶) and a large number of tissues (≥ lg) and cells (>10⁷) have good separation effects. The simplicity of Trizoe reagent operation allows for simultaneous processing of multiple samples. All operations can be completed within one hour. The total RNA extracted by Trizoe can avoid contamination of DNA and proteins. Therefore, it can perform RNA blotting analysis, dot hybridization, poly (A)+selection, in vitro translation, RNA enzyme protection analysis, and molecular cloning. If used for PCR, it is recommended to use amplification grade DNase I to process the extracted total RNA when two primers are located within a single exon.

Trizoe reagent can promote the precipitation of various RNAs of different genera and molecular weight sizes. For example, RNA agarose gel electrophoresis extracted from rat liver and stained with ethidium bromide showed many discontinuous high molecular weight bands (mRNA and hnRNA components) between 7 kb and 15 kb, and two dominant ribosomes~5 kb (28S) and~2 kb (18S) low molecular weight RNAs between 0.1 and 0.3 kb (tRNA, 5S). When the extracted RNA was diluted with TE, its A260/A280 ratio was>1.8.

Application

Suitable for rapid extraction of total RNA, DNA, and proteins from various animal and plant tissues/cells.

Note

1.When the dosage of Trizoe is less than 2ml, it is recommended to use clean disposable polypropylene material test tubes.

2.When the amount of Trizoe used is large, glass test tubes (Corex) or polypropylene test tubes can be used, and pre tested to ensure that the test tube can withstand the centrifugal force of 12000×g after adding Trizoe and chloroform. Do not use test tubes with cracks or damage.

3.Carefully balance the test tube before centrifugation.

4.Before centrifugation, the glass tube mouth must be covered with aluminum foil and sealed with paraffin film, while the polypropylene tube must be tightly covered.

5.Separate RNA samples from a small amount of tissue (1-10mg) or cells (10²~10⁴): Add 800μl of Trizoe to the tissue or cells. After the sample is cracked, chloroform is added and the extraction operation in step 2 is carried out. Before precipitating RNA with isopropanol, 5-10μg of RNA free glycogen was added as a carrier for the water sample layer. To reduce its viscosity, use a 26 syringe to aspirate twice before adding chloroform to cut off genomic DNA. Glycogen will remain in the water sample layer and co precipitate with RNA. It will not inhibit the synthesis of the first chain of reverse transcription reaction or PCR until it is concentrated to 4mg/ml.

6.After homogenization and before adding chloroform, the sample can be stored at -60~-70°C for at least one month. RNA precipitation (step 4, RNA rinsing) can be stored in 75% ethanol for at least one week at 2-8°C and for at least one year at -5-20°C.

7.The desktop centrifuge can achieve a maximum centrifugal force of 2.600×g. If the centrifugal time is extended to 30-60 minutes, the operations in steps 2 and 3 can be met.

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