Soil and Fecal RNA Extraction Kit
Broad-spectrum application, powerfully removes humic acid.
One-click copy of information
Instruction manualPrice:
¥ 2600
Item Number:
400-107
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Specification
- 50T
- 100μg
隐藏域元素占位
- Product Description
- Composition
- Precautions
-
Product preservation
HAR Buffer 1 should be stored at 4°C; the remaining components may be stored at room temperature (15–25°C) with a shelf life of one year. Transport at room temperature.
Product Overview
This product is suitable for the extraction of total RNA from various soil and fecal samples. By combining silica‑membrane purification technology with an efficient humic acid removal protocol, this kit effectively eliminates inhibitory substances such as humic acids, yielding high‑quality RNA. The resulting RNA can be directly used in PCR, qPCR, and other downstream applications.
User-provided
Reagents: anhydrous ethanol, water-saturated phenol.
Equipment and consumables: benchtop high-speed centrifuge, vortex mixer, bead mill (recommended), and 2 mL and 1.5 mL RNase‑free centrifuge tubes.
Pre-experiment preparation
As indicated in the table below, dilute Washing Buffer 1 and Washing Buffer 2 with anhydrous ethanol, and store the diluted solutions at room temperature.
Component Amount of anhydrous ethanol added Washing Buffer 1 12 mL Washing Buffer 2 64 mL l HAR Buffer 2A is a white solid suspension; shake well before use and dispense as needed.
l When stored at low temperatures, the SRL buffer may precipitate. Before use, place the entire reagent bottle in a 55°C water bath until the precipitate is completely dissolved, then mix thoroughly before use.
l The CFS buffer contains low-toxicity organic compounds and is somewhat volatile; please store it in a dry, well-ventilated area.
Instructions for Use
1. Weigh 200 mg of glass bead 1 and 200 mg of glass bead 2 into 2-mL centrifuge tubes, then add 0.5 g of soil or fecal sample.
2. Add 500 μL of SRL buffer, 40 μL of HAR Buffer 1, 400 μL of water-saturated phenol (to be prepared separately), and 400 μL of CFS buffer to each sample in a centrifuge tube, then vortex at high speed for 10 minutes. For better results, we recommend using a bead‑mill homogenizer, such as the FastPrep‑24.
3. Centrifuge at 13,000 g for 5 min at room temperature.
4. Add 70 μL of HAR Buffer 2A and 70 μL of HAR Buffer 2B, respectively, to a new 2 mL centrifuge tube, then add 350 μL of the supernatant, vortex to mix thoroughly, and incubate on ice for 3 minutes.
5. Centrifuge at 13,000 g for 2 min at room temperature, then carefully transfer 350 μL of the supernatant to a new 2 mL centrifuge tube.
6. Add an equal volume of DNA Binding Buffer and vortex to mix thoroughly.
7. Place the DNA adsorption column into the collection tube, transfer the mixture from step 6 into the binding column, and centrifuge at room temperature at 13,000 × g for 30 seconds. Discard the binding column and retain the filtrate.
8. Add an equal volume of RNA Binding Buffer to the filtrate and gently pipette up and down to mix.
9. Place the RNA binding column into a new collection tube, transfer 700 μL of the mixture from step 8 into the binding column, and centrifuge at 13,000 × g at room temperature for 30 seconds; discard the flow-through.
10. Repeat step 9 until all the mixture from step 8 has passed through the binding column.
11. Place the RNA binding column into the collection tube, add 500 μL of Washing Buffer 1 to the column, centrifuge at 13,000 g for 30 seconds, and discard the flow-through.
12. Place the RNA binding column into the collection tube, add 700 μL of Washing Buffer 2 to the binding column, centrifuge at 13,000 g for 30 seconds, and discard the supernatant.
13. Repeat step 12.
14. Place the RNA adsorption column into the collection tube and centrifuge at 13,000 × g for 2 minutes. Discard the supernatant.
15. Place the RNA binding column into a new 1.5 mL centrifuge tube, add 30–50 μL of Elution Buffer to the column, incubate at room temperature for 1–2 minutes, and then centrifuge at 13,000 × g for 1 minute to elute the RNA.
16. Store RNA at −80°C.
Frequently Asked Questions and Solutions
1. Low RNA yield
a. Low RNA content in samples: RNA levels vary considerably among different soil samples. For samples with low RNA content, the sample volume may be appropriately increased.
b. Inadequate sample lysis: Insufficient grinding; it is recommended to use a bead mill for grinding. If using a vortex mixer, set the speed to maximum. Excessive sample input can also lead to incomplete lysis; adjust by reducing the sample volume or proportionally increasing the amount of lysis buffer.
c. Washing Buffer 1 and Washing Buffer 2 were not supplemented with anhydrous ethanol as required: Please verify that anhydrous ethanol has been added prior to use.
2. RNA degradation
a. Excessive sample volume: Appropriately reduce the amount of sample loaded, or proportionally increase the volume of lysis buffer.
b. RNase contamination: Prior to use, treat the work surface with an RNase‑removal agent, and employ RNase‑free consumables and laboratoryware.
3. Low RNA purity
a. Humic acid contamination: Lower the ambient temperature during sample lysis to minimize humic acid release; increase the volume of HAR buffer 2A/2B used; however, both of these measures will to some extent reduce RNA yield.
b. Salt contamination: After adding Washing Buffer 1, let it stand for 5 minutes, then centrifuge.
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Product Composition
Item number Component Specifications 400-107A SRL buffer 28 mL 400-107B CFS buffer 22 mL 400-107C HAR Buffer 1 2.2 mL 400-107D HAR Buffer 2A 4 mL 400-107E HAR Buffer 2B 4 mL 400-107F DNA Binding Buffer 20 mL 400-107G RNA Binding Buffer 39 mL 400-107H Washing Buffer 1 18 mL 400-107I Washing Buffer 2 16mL 400-107J Elution Buffer 3 mL 400-107K gDNA Removal Column (with Collection Tube) 50 sets 400-107L RNA-binding column (with collection tube) 50 sets 400-107M Glass bead 1 14 g 400-107N Glass beads 2 14 g -
Precautions
1. Before first use, please add the specified amount of anhydrous ethanol to Washing Buffer 1 and Washing Buffer 2, respectively. After adding, promptly check the box to indicate that ethanol has been added, to avoid adding it multiple times.
2. When using this product, be sure to take appropriate personal protective measures, such as protective clothing, gloves, goggles, and a face shield.
3. This kit is intended for research use only.
Keywords
GOONIE
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Goonie Biotechnology (Guangzhou) Co., Ltd.
Room 321–332, Building G5, No. 39 Ruihe Road, Huangpu District, Guangzhou
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