Polysaccharide and Polyphenol Plant Total RNA (Più) Small Quantity Extraction Kit

1. Componenti del kit di reagenti

Specifiche	50T	100T
Gatto. NO.	SN0305AD	SN0306AD
RNA Extraction Columns (impostato)	50 (impostato)	100 (impostato)
DNA Clean-Up Columns (impostato)	50 (impostato)	100 (impostato)
RNA Extraction Buffer I	30 ml	2×30 ml
Inhibitor Removal Buffer	30 ml	2×30 ml
Tampone di lavaggio 1	15 ml	2×15 ml
Tampone di eluizione	20 ml	20 ml
Manuale di istruzioni	1	1

 

2. Magazzinaggio

This reagent kit can be stored at room temperature (15-25℃) in a dry environment and is stable for 12 mesi.

3. Istruzioni per l'uso del kit di reagenti

3.1 Questo kit è destinato a scopi di ricerca in biologia molecolare e non deve essere utilizzato per la diagnosi o il trattamento di malattie.

3.2 Alcuni componenti del kit contengono sostanze irritanti; è opportuno prendere le dovute precauzioni (come indossare indumenti protettivi e occhiali protettivi).

3.3 L'utilizzo di questo kit richiede apparecchiature aggiuntive come una centrifuga ad alta velocità, bagnomaria (bagno di metallo), miscelatore a vortice, etanolo anidro, azoto liquido, cloroformio, acqua deionizzata sterile, e tubi EP.

4. Introduzione al kit di reagenti

This RNA purification kit provides a fast and efficient method for purifying polysaccharide and polyphenol plant total RNA, suitable for most polysaccharide and polyphenol-rich plant tissues. In general, plant tissues rich in polysaccharides and polyphenols contain a high level of secondary phenolic metabolites and polysaccharides, significantly affecting RNA extraction efficiency. This kit can effectively remove polysaccharides and polyphenols, as well as efficiently eliminate DNA contamination from samples. If the experiment is sensitive to DNA, it is recommended to use primers spanning introns for downstream experiments.

The RNA Fast Purification Kit can extract total RNA from plant tissues (including nuclear RNA and cytoplasmic RNA) entro 1 ora. The extracted RNA can be directly used for RT-PCR, Northern blotting, eccetera. L'intero processo di purificazione non richiede reagenti tossici come il fenolo-cloroformio, making this RNA purification kit suitable for various other sample types.

5. Principi e procedure sperimentali

6. Processo di estrazione

Precauzioni prima di iniziare l'esperimento:

UN. Prima di usare Tampone di lavaggio 1, add the specified amount of absolute ethanol according to the label on the reagent bottle, and check the box on the label to indicate that absolute ethanol has been added.

B. Il buffer di eluizione è un 0.1x soluzione TE contenente una quantità minima di EDTA. Se l'EDTA influisce sugli esperimenti successivi, it is recommended to replace the Elution Buffer with sterile deionized water.

1. Elaborazione del campione:

UN. Raccolta e stoccaggio dei materiali:

If fresh materials cannot be used immediately, place them in liquid nitrogen and store them at -80°C.

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 di materiale secco utilizzando azoto liquido.

3. Aggiungere 600μl RNA Extraction Buffer I, ensuring there are no blocky tissues in the ground sample. Blocky tissues are difficult to lyse and can reduce RNA yield.

4. Vortex for 30 secondi.

5. Centrifugare il lisato 5 minuti a 12,000 giri/min.

(Nota: Polysaccharide plant materials may produce a lot of sticky substances at this step, which can shear RNA in subsequent steps. Perciò, remove these substances during this step.)

6. Transfer the supernatant obtained in the previous step to a DNA Clear Purification Column, centrifugare a 12,000 giri al minuto per 30 secondi, and collect the filtrate (Nota: RNA is present in the filtrate).
7. Aggiungere 250μl of absolute ethanol, mescolare pipettando. If there is a small amount of precipitation, it does not affect subsequent experiments. Add the liquid to the RNA purification column, centrifugare a 12,000 giri al minuto per 30 secondi, and discard the flow-through.
8. Aggiungere 700μl Inhibitor Removal Buffer to the RNA purification column, centrifugare a 12,000 giri al minuto per 30 secondi, and discard the flow-through.
9. Aggiungere 700μl Wash Buffer 1 to the RNA purification column, centrifugare a 12,000 giri al minuto per 30 secondi, and discard the flow-through.
10. Aggiungere 500μl Wash Buffer 1 to the RNA purification column, centrifugare a 12,000 giri al minuto per 3 minuti, and discard the flow-through.
11. Place the RNA purification column into a new 1.5ml centrifuge tube, air-dry the membrane at room temperature for 2 minuti.

(Nota: Confirm the addition of ethanol to Wash Buffer 1. The presence of ethanol significantly affects subsequent experiments. Perciò, membrane drying is crucial. Dopo la centrifugazione, ensure the absence of ethanol before elution. Eliminare i rifiuti e il tubo di raccolta. After using Wash Buffer 1, the membrane on the RNA purification column should only have a slight color. Carefully remove the RNA purification column after centrifugation, ensuring it does not touch the collection tube to avoid ethanol contamination.)

12. Gocciolare 50-100μl Elution Buffer sulla membrana, centrifugare a 12,000 giri al minuto per 1 minuto, and collect the RNA solution.

(Nota: Eluting RNA with 50μl Elution Buffer can increase RNA concentration but decrease total RNA yield.)