Manual

Product Number: RNK4001

Shipping and Storage

1. All solutions should be clear. If the ambient temperature is low and the solution may form a precipitate, it should not be used directly. It can be heated in a 37℃ water bath for a few minutes to restore clarity.
2. Inappropriate storage at low temperatures (4℃ or -20℃) can cause solution precipitation, affecting the effectiveness of use. Therefore, transportation and storage are carried out at room temperature (15℃ -25℃).
3. Wash Solution 2/3 may precipitate crystals after adding ethanol and using for a few days, but it does not affect its use. Simply do not absorb the crystals and absorb the supernatant before use.
4. To avoid the volatilization, oxidation, and pH changes of reagents exposed to air for a long time, the lid of each solution should be promptly closed after use.

Components

Component	Storage	RNK4001 50 Preps
Buffer RLT Plus	RT	50mL
PLANTaid	RT	5mL
Wash Solution 1	RT	12mL
		Add 28mL anhydrous ethanol before first use
Wash Solution 2/3	RT	10mL
		Add 42mL anhydrous ethanol before first use
RNase-free H2O	RT	5mL
Genomic DNA clearance column and collection tube	RT	50
RNase free adsorption column RA and collection tube	RT	50

Note: This reagent kit can be stored at room temperature for 12 months without affecting its effectiveness.

Description

Traditional microRNA extraction kits are made using the principle of guanidine isothiocyanate/phenol/chloroform (TRIzol method) combined with high concentration ethanol silica gel membrane to adsorb small fragments of microRNA. However, for complex plant species such as cotton, grapes, Populus euphratica, holly, roses, dendrobium, ginseng, rice seeds, ginseng, corn germ, and many other samples, the TRIzol principle cannot even extract the required total RNA, let alone microRNA. Therefore, world-renowned companies such as Qiagen and Invitrogen, which use the Trizol method principle for microRNA extraction kits, are helpless when faced with complex plant samples, and there are simply no complex plant microRNA extraction kits in their product lines. Users have no choice but to use traditional methods or TRIzol extraction, and isopropanol/ethanol precipitation method to extract microRNAs. Although some microRNAs can also be extracted, the precipitation method incurs huge losses or co precipitates with impurities, affecting experimental results. When submitting to international journals, they often face doubts and even have their papers revoked. On the basis of the company's global experience in extracting hundreds of complex plants, this kit innovatively uses a technology route that does not require phenol or chloroform to solve this problem. It can extract microRNAs from most complex plants such as cotton, poplar, holly, rose, and Danshen. Of course, this kit is also suitable for various simple samples such as Arabidopsis, rice, wheat, tobacco, and rice.

The unique lysis buffer rapidly lyses cells and inactivates cell RNA enzymes. Plant RNA enhancer PLANTaid helps bind to polysaccharides and polyphenols, which are removed by centrifugation. The lysis mixture is then passed through a genomic DNA clearance column, where genomic DNA is cleared and RNA (including microRNA) is selectively filtered. After adjusting the binding conditions of the filtered RNA with ethanol, the RNA selectively adsorbs onto the silica membrane inside the centrifuge column in a high salt state. Then, a series of special rinsing centrifugation steps are performed to quickly remove cellular metabolites, proteins, and other impurities. Finally, low salt RNase free H20 elutes pure RNA (including microRNA) from the silica membrane.

The separation and enrichment of microRNA components and total RNA (>200 nt) can also be obtained separately by using the extraction operation steps.

Application

Suitable for rapid extraction of plant microRNA or separate extraction of microRNA/total RNA.

Features

1. Completely avoid using toxic reagents such as phenol and chloroform, and do not require steps such as ethanol precipitation.
2. Fast, simple, and the operation of a single sample can generally be completed within 30 minutes.
3. The unique plant RNA enhancer can effectively bind to polysaccharide polyphenols and improve clearance efficiency.
4. Multiple column rinses ensure high purity, with a typical OD₂₆₀/OD₂₈₀ ratio of 2.0-2.2, which can be used for RT-PCR, Northern blot, and various experiments.

---