Simple Plant Tissue RNA Extraction Kit
11-minute rapid extraction, 0 phenol, 0 chloroform
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Instruction manual- Product Description
- Composition
- Precautions
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Product Storage
Store and transport at room temperature (15–25°C); shelf life is one year.
Product Overview
This kit is suitable for the rapid extraction of total RNA from simple plant tissues. It can be used to isolate RNA from a variety of common plant tissues, including young tissues of rice, wheat, maize, Arabidopsis, tobacco, and others. During the extraction process, this kit does not require the use of toxic reagents such as phenol–chloroform or β‑mercaptoethanol.
There is no need for time‑consuming alcohol precipitation; the entire extraction process takes only 11 minutes. The kit’s specialized gDNA‑removal column effectively eliminates contaminants and genomic DNA, while the RNA‑binding column efficiently captures RNA. Coupled with an optimized buffer, this system yields high‑purity total RNA with minimal residual gDNA and no contamination from proteins or other impurities. The resulting RNA is suitable for a wide range of downstream applications, including RT‑PCR, quantitative real‑time PCR, RNA library construction, microarray analysis, Northern blotting, dot blotting, and molecular cloning.
Pre-experiment preparations
1. Before first use, 20 mL of anhydrous ethanol must be added to the CB Buffer and thoroughly shaken.
2. Before first use, add 48 mL of anhydrous ethanol to the Wash Buffer and shake thoroughly.
3. For first-time use, it is recommended to aliquot the Elution Buffer into small portions in a clean bench.
Instructions for Use
1. Sample Processing
Weigh 50–100 mg of plant tissue, add liquid nitrogen, and grind it into a powder using a mortar, or rapidly freeze the sample in liquid nitrogen and then use a cryogenic grinder to quickly pulverize it. The grinding should be as thorough and rapid as possible, while maintaining low temperatures to prevent RNA degradation.
2. Sample Lysis
Quickly add 600 μL of Lysis Buffer to the finely ground plant sample powder, vortex vigorously for 30 seconds to ensure thorough mixing, then centrifuge at 13,000 × g at room temperature for 5 minutes.
3. gDNA removal
Discard the pellet, and immediately transfer the supernatant from the previous centrifugation step into the gDNA removal column (which has been pre‑assembled in the collection tube). Centrifuge at 13,000 × g at room temperature for 30 seconds. The product remains in the collection tube; retain the filtrate in the tube.
4. RNA Binding
Add anhydrous ethanol to the collection tube at a volume equal to 0.5 times the volume of the eluate, pipette up and down 6–8 times to mix thoroughly, then transfer the mixture into the RNA-binding column (with the column first placed inside the collection tube). Centrifuge at 13,000 × g at room temperature for 30 seconds.
5. Column Cleaning
1) Discard the filtrate, add 700 μL of CB Buffer (add the corresponding amount of anhydrous ethanol before first use) to the RNA binding column (including the collection tube), and centrifuge at 13,000 × g at room temperature for 30 seconds.
2) Discard the filtrate, add 500 μL of Wash Buffer to the RNA-binding column (including the collection tube)—prior to first use, add the corresponding volume of anhydrous ethanol—and centrifuge at 13,000 × g at room temperature for 30 seconds.
3) Repeat step 2 above once.
4) Discard the filtrate, and place the RNA-binding column back into the collection tube. Centrifuge at 13,000 × g at room temperature for 1 minute.
6. RNA Elution
1) Discard the collection tube, transfer the RNA-binding column into a new 1.5 mL RNase-free centrifuge tube, and leave it open under the laminar flow hood at maximum fan speed for 1 minute to dry.
2) Add 30–100 μL of Elution Buffer dropwise to the center of the column membrane, then centrifuge at 13,000 × g at room temperature for 1 minute. The product in the 1.5 mL centrifuge tube is the RNA, which can be used immediately for downstream applications or stored at −80°C.
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Product Composition
Product Number Component (50 times) 400-106A Lysis Buffer 40 mL 400-106B CB Buffer 20 mL 400-106C Wash Buffer 12 mL 400-106D Elution Buffer 10 mL 400-106E gDNA Removal Column (with Collection Tube) 50 sets 400-106F RNA-binding column (with collection tube) 50 sets -
Precautions
1. Please use high-quality centrifuge tubes to prevent tube breakage or reagent spillage during the experiment.
2. To ensure extraction quality, it is recommended to use RNase‑free reagents and consumables throughout the entire process, particularly during the RNA elution step.
3. For rare samples or difficult-to-extract samples whose extraction efficiency is uncertain, please conduct a preliminary experiment using a small number of samples before the formal experiment to evaluate the extraction performance.
Keywords
GOONIE
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Goonie Biotechnology (Guangzhou) Co., Ltd.
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