Transport temperature:Normal temperature transportation

Product content:

 

Content	50T	100T	Save	Validity period
Lysis solution	50ml	100ml	2-8℃	One ​​year
Rinse	15ml	30ml	RT	One ​​year
Column washing solution	50ml	100ml	RT	One ​​year
RNase free ddH2O	15ml	30ml	RT	One ​​year
RNase free adsorption column	50	100	RT	One ​​year
RNase free collection tube (2ml)	50	100	RT	One ​​year

Operation steps:1. Sample processing: a. Plant tissue: Take 100 mg of tissue fresh or frozen at -70°C and grind it in liquid nitrogen. Add the powder to 1 ml of lysis solution and mix well.

b. Animal tissue: Take 100mg of fresh or -70℃ frozen tissue, add 1ml of lysis solution, and use a tissue grinding pestle or homogenizer Homogenization treatment.

c. Adherent cells: add lysis solution directly to the culture plate to lyse the cells, every 10⁶ cells Add 1ml of lysis buffer. Use a sampler to mix evenly.

d.Cell suspension: centrifuge to collect cells. Add 1 ml of lysis buffer for every 10⁶ animal, plant and yeast cells or every 10⁷ bacterial cells and mix well.

e. Blood processing: Take 0.2-1ml of fresh blood and add 3 times the volume of red blood cell lysate, mix and place at room temperature for 10 minutes, 10000rpm Centrifuge for 1 minute. Discard the supernatant. If the pellet contains red blood cells, add 2 times the volume of red blood cell lysis buffer and repeat the lysis step. After centrifugation, add 1 ml of lysis buffer to the precipitate and mix well.

2. Place the processed sample at room temperature for 5 minutes to completely separate the nucleic acid-protein complexes.

3. Add 0.2ml chloroform to the homogenized sample, cover the tube cap, shake vigorously for 15 seconds, and leave it at room temperature for 3-5 minutes .

4. Centrifuge at 12000 rpm at 2-8℃ for 10 minutes. RNA is mainly in the upper colorless aqueous phase. Transfer the aqueous phase to a new tube and do not absorb the precipitate.

5. Adsorption column pre-treatment: add 500ul column washing solution to the adsorption column, leave it at room temperature for 2 minutes, 2-8℃ 12,000 rpm Centrifuge for 2 minutes and discard the waste liquid.

6. Add 200ul of absolute ethanol to the supernatant collected in step 4, mix well, add to the adsorption column and let stand for 2 minutes, 2- Centrifuge at 8°C 12000rpm for 2 minutes and discard the waste liquid.

7. Add 600ul of rinse solution to the adsorption column (please check whether absolute ethanol has been added before use), 2-8℃ Centrifuge at 12,000 rpm for 2 minutes and discard the waste liquid.

8. Add 600ul of rinse solution to the adsorption column, centrifuge at 12,000 rpm at 2-8°C for 2 minutes, and discard the waste liquid.

9. Centrifuge at 12000rpm for 2 minutes, discard the collection tube, and place the adsorption column at room temperature for a few minutes to remove the residual rinse fluid in the adsorption column.

10. Put the adsorption column into a new tube, drop 50-100ul RNase free ddH2O into the center of the membrane, leave it at room temperature for 5 minutes, and centrifuge at 12000rpm for 2 minutes at room temperature to obtain RNA.

Notes:

 

1. All related utensils and consumables should be RNase-free products. Be careful during the operation and wear masks and gloves to avoid contaminating samples with RNase in the environment.

2. The OD value of RNA in aqueous solution may be between 1.5-1.9, but this does not mean that the RNA is impure and needs electrophoresis detection.

 

Remarks:

The above data are all from public literature and have not been independently verified by Sunlong Biotech. They are for reference only.

These protocols are for reference only. Sunlong Biotech does not independently validate these methods.

 

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