- Bestandteile des Reagenzienkits
| Spezifikationen | 50T | 100T |
| Katze. NEIN. | SN0305PD | SN0306PD |
| RNA -Extraktionssäulen (Satz) | 50 (Satz) | 100 (Satz) |
| DNA Clean-Up Columns (Satz) | 50 (Satz) | 100 (Satz) |
| Inhibitor Removal Purification Columns (Satz) | 50 (Satz) | 100 (Satz) |
| RNA Extraction Buffer I | 30 ml | 2×30 ml |
| Inhibitorentfernungspuffer | 30 ml | 2×30 ml |
| Waschpuffer 1 | 15 ml | 2×15 ml |
| Elutionspuffer | 20 ml | 20 ml |
| Bedienungsanleitung | 1 | 1 |
- Lagerung
This reagent kit can be stored at room temperature (15-25℃) in einer trockenen Umgebung und ist stabil für 12 Monate.
- Anweisungen zur Verwendung des Reagenzienkits
3.1 Dieses Kit ist für molekularbiologische Forschungszwecke bestimmt und sollte nicht zur Diagnose oder Behandlung von Krankheiten verwendet werden.
3.2 Einige Bestandteile des Kits enthalten Reizstoffe; Es ist ratsam, die erforderlichen Vorsichtsmaßnahmen zu treffen (wie das Tragen von Schutzkleidung und Schutzbrillen).
3.3 Die Verwendung dieses Kits erfordert zusätzliche Ausrüstung wie eine Hochgeschwindigkeitszentrifuge, Wasserbad (Metallbad), Vortexmixer, Wasserfreies Ethanol, flüssiger Stickstoff, Chloroform, steriles entionisiertes Wasser, und EP-Röhren.
- Einführung in das Reagenzienkit
This RNA purification reagent kit provides a fast and effective purification of plant total RNA containing polysaccharides, Lipide, polyphenols, and other components. It is suitable for most complex plant tissues. In general, lipid-rich plant tissues contain a high content of lipid compounds, which significantly affect RNA extraction efficiency. This kit utilizes exclusive inhibitor removal columns to effectively eliminate lipids from plant samples, as well as to remove DNA contamination. If the experiment is sensitive to DNA, it is recommended to use intron-spanning primers for downstream experiments.
The RNA rapid purification reagent kit can extract plant total RNA (einschließlich nuklearer RNA und zytoplasmatischer RNA) innerhalb 1 Stunde. Die extrahierte RNA kann direkt für RT verwendet werden-PCR, Northern Blot, usw. The entire purification process does not require toxic reagents such as chloroform, making the RNA purification reagent kit suitable for various other samples.
- Experimentelle Prinzipien und Verfahren
- Extraktionsprozess
Vorsichtsmaßnahmen vor Beginn des Experiments:
A. Waschen Puffer 1: Prior to use, add the specified amount of absolute ethanol as indicated on the reagent bottle label. Check the label to confirm the addition of absolute ethanol.
B. Elutionspuffer ist ein 0.1x TE-Lösung with a minimal amount of EDTA. If EDTA may affect subsequent experiments, it is recommended to replace the elution buffer with sterile deionized water.
C. RNA Extraction Buffer I contains a small amount of phenol, which may precipitate. Vor Gebrauch, mix thoroughly by warming in a water bath; after use, store away from light.
- Probenverarbeitung:
A. Materialsammlung und -lagerung:
If freshly collected materials cannot be immediately used, place them in liquid nitrogen and store them at -80℃.
B. Whenever possible, collect fresh materials as they contain fewer polysaccharides and polyphenols.
2. Grind approximately 100 mg of fresh samples or not more than 20 mg of dry material with liquid nitrogen.
3. Hinzufügen 600μl of RNA Extraction Buffer I, ensuring there are no tissue clumps in the ground sample. Tissue clumps are difficult to lyse and can reduce RNA yield.
4. Vortex for 30 S.
5. Transfer the lysate to aninhibitor removal purification column, Zentrifuge bei 12,000 U/min für 5 Mindest.
(Notiz: Lipid-rich plant materials may contain many lipid compounds at this step, which can affect RNA extraction. Remove these substances during this step. If buffer residue remains in the inhibitor removal purification column during centrifugation, extend centrifugation time appropriately.)
- Transfer the obtained supernatant to a DNA removal column, Zentrifuge bei 12,000 rpm for 30s, and collect the filtrate (Notiz: RNA is present in the filtrate).
- Hinzufügen 250μl von absolutem Ethanol, Mischen Sie durch Pipettieren. If there is a small amount of precipitation, it does not affect subsequent experiments. Transfer the liquid to an RNA purification column, Zentrifuge bei 12,000 rpm for 30s, discard the flow-through.
- Hinzufügen 700μl of inhibitor removal buffer, Zentrifuge bei 12,000 rpm for 30s, discard the flow-through.
- Hinzufügen 700μl von waschenbuffer 1 zur RNA -Reinigungsspalte, Zentrifuge bei 12,000 rpm for 30s, discard the flow-through.
- Hinzufügen 500μl von waschen buffer 1 zur RNA -Reinigungsspalte, Zentrifuge bei 12,000 U/min für 3 Mindest, discard the flow-through.
- Place the RNA purification column into a new 1.5ml centrifuge tube, air-dry the membrane at room temperature for 2 Protokoll.
(Notiz: Confirm that absolute ethanol has been added to waschen buffer 1. The presence of ethanol has a serious impact on subsequent experiments, so drying the membrane is crucial. Nach Zentrifugation, ensure no ethanol is present before elution, then discard the waste and collection tube. After washing with wash buffer 1, the membrane on the RNA purification column should have only a slight color. Nach Zentrifugation, carefully remove the RNA purification column, ensuring it does not touch the collection tube to avoid ethanol interference.)
- Aerially pipette 50-100μl Elutionspuffer auf die Membran, Zentrifuge bei 12,000 U/min für 1 Mindest, and collect the RNA solution.
(Notiz: Elusionsrna mit 50 μl of elution buffer can increase RNA concentration but reduces total RNA yield.)
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