RNAplant plant RNA extraction reagent instructions

Plant RNA extraction reagent

Product introduction:

RNAplant reagents extract high-purity total RNA from plant tissues, especially plant tissues rich in polyphenols or starches (eg potato tubers, white pine needles or tender shoots, and tomato leaves, etc.).

200 mg of tissue can be processed 200 times per 100 ml.

Storage conditions: transport at room temperature, stored at 4 ° C, can be stored for 6-12 months.

Prepare reagents : liquid nitrogen, mortar, RNase-free centrifuge tube, 5 M NaCl ( no RNase water ), chloroform, isopropanol, 75% ethanol (no RNase water), no RNase water.

Small-scale extraction procedure : (sample < 0.1 g , using small-scale extraction method):

1. Take no more than 0.1 g of plant tissue, grind it in liquid nitrogen, transfer to a 1.5 ml centrifuge tube, add 0.5 ml of extraction reagent, and shake until thoroughly mixed.

Note: Do not irritate too much, otherwise it will easily lead to genomic DNA contamination.

2. Leave at room temperature for 5 minutes.

3. Centrifuge at 12, 000 rpm for 2-3 minutes at 4 ° C. Transfer the supernatant to a new RNase-free centrifuge tube.

4. Add 0.1 ml of 5 M NaCl ( prepared without RNase water ) to the supernatant and mix gently.

5. Add 0.3 ml of chloroform and mix upside down.

6. Centrifuge at 12,000 rpm for 10-15 minutes at 4 ° C. Transfer the upper aqueous phase to a new RNase-free centrifuge tube.

7. Add equal volume of isopropanol to the resulting water, mix and let stand for 10 minutes at room temperature.

8. Centrifuge at 12,000 rpm for 10-15 minutes at 4 °C. Discard the supernatant and take care not to pour out the sediment. Rinse the pellet with 1 ml of 75% ethanol. (Precipitation may be difficult to see and should be handled with care.)

9. Centrifuge at 5000-7000 rpm for 3 minutes at 4 °C. Pour out the liquid, taking care not to pour out the sediment. The remaining small amount of liquid was briefly centrifuged, sucked out with a pipette tip, and allowed to air dry for 1-2 minutes at room temperature until the precipitate was dry.

10. Add appropriate amount of RNase-free water to fully dissolve RNA and store at -70 °C.

Precautions:

RNAplant has a pungent taste and should be done in a well ventilated place during operation.