ARN total de planta compleja (más) Kit de extracción de cantidad pequeña

1. Componentes del kit de reactivos

Especificaciones	50t	100t
Gato. No.	SN0305PD	SN0306PD
Columnas de extracción de ARN (colocar)	50 (colocar)	100 (colocar)
Columnas de limpieza de ADN (colocar)	50 (colocar)	100 (colocar)
Inhibitor Removal Purification Columns (colocar)	50 (colocar)	100 (colocar)
Extracción de ARN Buffer I	30 ml	2×30 ml
Tampón de eliminación de inhibidores	30 ml	2×30 ml
Tampón de lavado 1	15 ml	2×15 ml
Tampón de elución	20 ml	20 ml
Manual de instrucciones	1	1

2. Almacenamiento

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

3. Instrucciones para usar el kit de reactivos

3.1 Este kit está destinado a fines de investigación de biología molecular y no debe utilizarse para el diagnóstico o tratamiento de enfermedades..

3.2 Algunos componentes del kit contienen irritantes.; es recomendable tomar las precauciones necesarias (como usar ropa y gafas protectoras).

3.3 El uso de este kit requiere equipo adicional como una centrífuga de alta velocidad., baño de agua (baño de metal), mezclador Vortex, etanol anhidro, nitrógeno líquido, cloroformo, agua desionizada estéril, y tubos EP.

4. Introducción al kit de reactivos

This RNA purification reagent kit provides a fast and effective purification of plant total RNA containing polysaccharides, lípidos, 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 (Incluyendo ARN nuclear y ARN citoplasmático.) dentro 1 hora. The extracted RNA can be directly used for RT-PCR, transferencia Northern, etc.. The entire purification process does not require toxic reagents such as chloroform, making the RNA purification reagent kit suitable for various other samples.

5. Principios y procedimientos experimentales

6. Proceso de extracción

Precauciones antes de comenzar el experimento.:

A. Lavar Buffer 1: Antes de usar, 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. El tampón de elución es un 0.1x solución TE 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. Antes de usar, mix thoroughly by warming in a water bath; after use, store away from light.

 

1. Procesamiento de muestras:

A. Recolección y almacenamiento de materiales:

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. Agregar 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, centrifugar en 12,000 rpm para 5 mín..

(Nota: 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.)

6. Transfer the obtained supernatant to a DNA removal column, centrifugar en 12,000 rpm for 30s, and collect the filtrate (Nota: RNA is present in the filtrate).
7. Agregar 250μl of absolute ethanol, mezclar pipeteando. If there is a small amount of precipitation, it does not affect subsequent experiments. Transfer the liquid to an RNA purification column, centrifugar en 12,000 rpm for 30s, discard the flow-through.
8. Agregar 700μl of inhibitor removal buffer, centrifugar en 12,000 rpm for 30s, discard the flow-through.
9. Agregar 700µl de lavarbuffer 1 a la columna de purificación de ARN, centrifugar en 12,000 rpm for 30s, discard the flow-through.
10. Agregar 500µl de lavar buffer 1 a la columna de purificación de ARN, centrifugar en 12,000 rpm para 3 mín., 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 minutos.

(Nota: Confirm that absolute ethanol has been added to lavar buffer 1. The presence of ethanol has a serious impact on subsequent experiments, so drying the membrane is crucial. Después de la centrifugación, ensure no ethanol is present before elution, luego deseche los residuos y el tubo de recolección.. After washing with wash buffer 1, the membrane on the RNA purification column should have only a slight color. Después de la centrifugación, Retire con cuidado la columna de purificación de ARN., ensuring it does not touch the collection tube to avoid ethanol interference.)

12. Aerially pipette 50-100µl de tampón de elución sobre la membrana, centrifugar en 12,000 rpm para 1 mín., and collect the RNA solution.

(Nota: Eluting RNA with 50 μl of elution buffer can increase RNA concentration but reduces total RNA yield.)