I am working on RNA extraction of pineapple fruits, however my fruit materials are in another country and are therefore not very accessible to me. I need to test different methods for RNA extraction on the fruits (CTAB vs. Trizol vs. kits, etc.).
My question is: would fruits from the grocery store still have good quality RNA in them? After being removed from the plant, shipped, and sitting in the grocery store for a few days, the fruit would still be alive of course, and the RNA expression profile would change, but I just need to know that it is possible to get RNA that is good enough quality for RT-PCR and subsequent NGS library prep from store-bought fruit. Would the RNA start to degrade when the fruit is harvested, or would it not begin to degrade until the flesh is excised from the fruit?
Eventually, I will be receiving fruit tissue at all different development stages, but at this point I am just trying to find a protocol that will work for giving me good quality (RIN of 7.5 or higher) RNA from the fruit tissue, and grocery store fruits are the most accessible/replaceable to me at the moment.
Looking at this paper:
https://www.nature.com/articles/s41598-017-14239-6
as long as your technique is good, and you have RNA handling experience, it looks like you should have no difficulty in obtaining RNA-seq quality samples from fruit, post harvest.
you can use chloroform-phenol extraction method but, in lowering ph to 4 .it easy and rapid method.
I would not worry, researchers have been buying from grocery stores and extracting DNA and RNA for ages, even from very sensitive types of tissues, such as spinach leaves. So, as long as your pineapple fruit is not already getting soft, indicating significant cell degradation, you should be able to easily find the right method for the job. And pineapples typically arrive in grocery stores in an unripe, relatively green and hard state. So, that will work to your advantage.
That being said, process the excised fruit flesh sample immediately and then store the RNA in a stabilizing solution (various are commercially available), if you do not progress to pure RNA right away. In the latter case, purified RNA samples should be stored at < -70°C. To avoid RNA degradation through repeated freezing/thawing, you should store them in aliquots that are only frozen and thawed once.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA