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Plant Seed RNA Extraction Kit(SpinColumn)

Packing Specification：

For research use only. Not for use in medicine, clinical diagnosis, food, cosmetics or other applications.

Product Introduction：
This is a rapid RNA extraction kit for plant seeds. Its unique working system enables rapid removal of starch, polysaccharides, fats, proteins, and other components from plant seeds through simple operations, releasing RNA. It can be used in various molecular biology experiments such as RT-PCR, Real-Time PCR, Northern blot, Dot blot, poly A selection, in vitro translation, and cDNA library construction.

Product Features：
1.No toxic reagents such as phenol or chloroform are used, and no ethanol precipitation steps are required.
2.Simple and fast: Operation for a single sample can generally be completed within 35 minutes, making it one of the simplest and fastest kits available.
3.Genomic DNA removal column technology effectively eliminates gDNA residue. The obtained RNA generally does not require DNase digestion and can be used in reverse transcription PCR (RT-PCR), quantitative real-time PCR, and other experiments.
4.Wide adaptability: Can extract RNA from various plant seeds.
5.Multiple column washing steps ensure high purity: The typical OD260/OD280 ratio ranges from 1.9 to 2.2, with almost no DNA residue. Suitable for RT-PCR, Northern blot, next-generation sequencing, and various other experiments.

Application Scope：
Suitable for the extraction of total RNA from plant seeds, such as wheat seeds, rice seeds, sesame seeds, dried corn seeds, fresh corn seeds, pea seeds, red bean seeds, mung bean seeds, sunflower seeds, watermelon seeds, etc.

Experimental Procedures：
Sample Pretreatment:Take 50-100 mg of plant seeds and grind them rapidly into powder in liquid nitrogen. Add 1 mL of Lysis Buffer SSB and vortex vigorously to mix well.Note: Most RNA in plant seeds is distributed in the embryo, so it is best to isolate the embryo separately if possible.
1.Vigorously shake the homogenized lysate for 15-20 seconds, then centrifuge at 13,000 rpm for 5-10 minutes to pellet impurities.

2.Transfer the supernatant to a new centrifuge tube, add 0.5 volumes of absolute ethanol (e.g., add 400 μL of absolute ethanol for 800 μL of supernatant), and immediately pipette to mix well.

3.Transfer the mixed solution (less than 720 μL each time, can be added in two portions) to a genomic DNA removal column, centrifuge at 13,000 rpm for 30 seconds, and discard the waste liquid.

4.Place the removal column into a new 2 mL centrifuge tube, add 500 μL of Lysis Buffer RLT Plus, centrifuge at 13,000 rpm for 30 seconds, and collect the filtrate (RNA is in the filtrate). Add 250 μL of absolute ethanol to the filtrate and immediately pipette to mix well.

5.Transfer the mixture from Step 5 to a SpinRA column, centrifuge at 13,000 rpm for 1 minute, and discard the waste liquid.

6.Add 700 μL of Buffer RW1, incubate at room temperature for 1 minute, centrifuge at 13,000 rpm for 30 seconds, and discard the waste liquid.

7.Add 700 μL of Buffer RW (please check if absolute ethanol has been added first!), centrifuge at 13,000 rpm for 30 seconds, and discard the waste liquid.

8.Add 500 μL of Buffer RW, centrifuge at 13,000 rpm for 3 minutes.

Carefully remove the SpinRA column (avoid the bottom of the SpinRA column touching the waste liquid in the collection tube), place it into a new RNase-free centrifuge tube. Add 30-50 μL of RNase-free H₂O to the center of the adsorption membrane, incubate at room temperature for 1 minute, and centrifuge at 12,000 rpm for 1 minute to obtain the RNA solution.