Hi dear colleagues, I work on altered expreeion profiles genes in bread wheat against mycotoxin of fusarium head blight.I need information about Sampling and RNA extraction from wheat spike. I'm looking for the best way to sampling from wheat spikes and RNA extraction problems.
Dear Montana
Greeting
Please find attached relevent information for you.This might be helpful to you.Good luck.
RNA extraction from developing or mature wheat seeds.
Furtado A. Methods Mol Biol. 2014.
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Abstract
Cereal grains, as storage tissues of the plant, contain high amounts of starch. Purification of RNA from plant tissue especially from seed tissue can be challenging due to this high starch content. Starch coprecipitates with RNA in the presence of isopropanol or ethanol and can interfere with the extraction process and downstream reactions. Thus the removal of starch by using appropriate methods is necessary for obtaining pure RNA to be processed for functional genomics analysis. We describe a method to isolate large amount of good-quality RNA from developing and mature wheat grain which can also be adapted to other cereal grains.
Link is
https://www.ncbi.nlm.nih.gov/pubmed/24243193/?ncbi_mmode=std
Dear Mojtaba
The main thing that you should take into consideration is the sampling time. In your case sampling should be carried out during infection phase (which is flowering phase in F. graminearum )
I really don't know which gene(s) you're studying, but I'm assuming that it should be expressed in this phase to form some sort of resistance. I suggest taking your samples during this period and flash freeze in field.
As far as I know, conventional RNA extraction methods should work fine with you, nothing to worry about.
Regards
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