Generally prior to RNA Extraction from plant, samples are centrifuged at 10K rpm for 5-10 mins at room temp followed up by one of the methods eg. Trizol method. And also, samples are always kept at - 20, no warmer temp req. as per my knowledge.

When working with RNA extraction from plant samples, it's important to ensure that the samples are properly preserved to prevent degradation. Generally, keeping the samples at a temperature below -80°C is recommended to maintain RNA integrity.

If you need to briefly thaw the samples for RNA extraction, it's important to work quickly to minimize RNA degradation. Thawing samples on ice or in a cold room can help slow down degradation processes. Keep in mind that RNA is sensitive to temperature changes, so minimize the time the samples spend at higher temperatures.

The exact time the samples can be kept warmer than -80°C will depend on various factors, including the specific plant species, the condition of the samples, and the downstream applications for the extracted RNA. As a general guideline, aim to keep the samples at warmer temperatures for the shortest time possible to avoid RNA degradation. If possible, perform the RNA extraction on ice or at a temperature close to 0°C to ensure RNA integrity. If you need to store the samples at warmer temperatures temporarily, it's best to do so for only a few minutes to an hour.

Ultimately, the goal is to minimize any temperature-induced RNA degradation during the extraction process. If you are unsure, you may want to consult established RNA extraction protocols specific to your plant species or seek advice from colleagues or experts experienced in RNA extraction from plant buds. Yetong Lu

You can put RNA in -20 without freez and thaw

It is better to combine two methods (trizole first two steps, use aqueous phase then proceed with the kit as per their instruction). therefore check the integrity using gel.

you will get better yield and good integrity.