RNA extraction from trout embryos
Posted 08 July 2010 - 03:17 AM
I've been having problems trying to extract RNA using TRI reagent from early stage trout embryos which have very small amounts of RNA and lots of lipid/protein. The problem seems to be that I get such small amounts of RNA that protein and phenol contamination becomes very evident causing very low 260/280 and 260/230 ratios. I've tried a high salt solution modifcation (NaCl and sodium citrate) of the isopropanol precipitation step and an additional ethanol/sodium acetate wash at the end of the protocol and both of these seem to remove some of the protein/phenol contamination, but also reduce my RNA yield further so the overall quality is still very bad.
If anyone has any ideas of anything else I can try that would be great!
Posted 08 July 2010 - 07:12 AM
Edited by mdfenko, 08 July 2010 - 07:12 AM.
genius does what it must
i do what i get paid to do
Posted 08 July 2010 - 11:02 AM
Posted 09 July 2010 - 03:19 AM
you can remove phenol with chloroform (or chloroform/isoamyl alcohol).
Thanks- I will give it a go.