I am working on a freshwater red algae. This algae is gelatinous in nature. I have been doing a series of extractions using various kits but couldn't get a good quality of RNA. Is there anyone who has worked on freshwater red algae and has done RNA extractions? Although the concentration is kind of okay (~40 ng/ul), the RIN score is always lower than 5. Any help, suggestions would be much appreciated. Thank you.
Adding sand while crushing the algae and using CTAB method can give a pure and more quantity of RNA as suggested by this paper.
I have also tried a CTAB method, but couldn't get a good RIN score. The problem with it, what I guess is, rapid RNA degradation during disruption. I have used 6 different types of kits so far and all of those worked for Arabidopsis but not for algae.
Catarina Figueiredo Mota
, Thank you for your suggestion. I don't let the sample thaw. As soon a I take the samples out of -80, I put them in a buffer for disruption using a bead beater. I also tried grinding in liquid N2, where I put tubes instantly in liquid N2 from -80 and don't let samples thaw or liquid N2 run out during grinding. As my sample is not a pure culture and is mucousy with a lot of polysaccharides, RNA is being degraded easily.- Badges
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