The samples are Prawns muscle and are stored in ethanol 100% for a day at RT and later stored at -20ºC (2 weeks)
I want to isolate RNA and DNA, just for a simple quantification and maybe later do RT-PCR.
I would verify your RNA integrity, once extracted (whichever protocol you choose). RNA is very labile, and even being stored at 4C for a day is certain to reduce the quality of your mRNA. Even storage at -20 is not ideal and can damage your samples; -80c would be much better. if you are looking to investigate relative transcription it may bias your results.
I'm not saying your samples will be bad (they may be fine), but you should definitely make sure before moving on to analysis, if only to strengthen your argument.
Usually in our RNA isolations we use ethanol to wash the pellet of RNA before saving in RNase free water. A kit might be useful, however there are protocols out there that do not require a kit.
Qiamp RNA blood mini kit permit to isolate RNA from 200 000 cells (leukocytes) or more. Ethanol induces nucleic acids precipitation and facilitates their fixation on silicat columns include in Qiamp kit. Use of Ethanol containing solution is necessary for wash nucleic acid pellets, avoiding their dissolution in washing solution.
I would verify your RNA integrity, once extracted (whichever protocol you choose). RNA is very labile, and even being stored at 4C for a day is certain to reduce the quality of your mRNA. Even storage at -20 is not ideal and can damage your samples; -80c would be much better. if you are looking to investigate relative transcription it may bias your results.
I'm not saying your samples will be bad (they may be fine), but you should definitely make sure before moving on to analysis, if only to strengthen your argument.
I think Enos c Kline is very true and I am also not sure the goodness of your samples stored in 100% ethanol.
If it is blood sample I suggest using PAXgene tubes to preserve your blood sample. The PAXgene tubes contain an RNA stabilizer that can stabilize intracellular RNA from whole blood for years if you freeze your sample after collection in -20C or -80 freezer. You can order Paxgene kit from Qiagen. The entire kit might be expensive. You can order only the PAXgene tube and use the preserved blood samples with ZR Whole-Blood RNA MiniPrep from Zymo Research. You need to modify the protocol little bit. We have a protocol that we use for years and we isolate high quality RNA from a frozen blood sample. If you need it email me I will send it to you.
Roche RNeasy kit is very easy to use and some of my samples (only different storage conditions), may have contained ethanol residues.
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