RNA Extraction using Trizol
Posted 21 January 2005 - 06:33 AM
Posted 21 January 2005 - 08:24 AM
Posted 21 January 2005 - 10:35 AM
Posted 21 January 2005 - 12:18 PM
Thanks for the help! One more thing...glycogen isn't in my protocol. How much glycogen should I use in the isopropyl precipitation step...and what does it do exactly?
With Trizol, I believe that the more quantity of the reagent you use, the better the results you get. I work with small tissue samples and always use 1 ml Trizol. Don't forget you need to add a carrier such as glycogen in the isopropyl precipitation step.
Posted 21 January 2005 - 03:17 PM
You can use 20 ug per sample. That amount will allow you to precipitate pg-amount of RNA from a volume of 1 ml.
Posted 24 January 2005 - 07:32 AM
Glycogen is a carrier for the precipitation of nucleic acids (DNA or RNA). It is not a must for RNA extraction with trizol.
Yeah, you don't need to add glycogen if your sample is big enough, but I found that glycogen is necessary when working with small samples of tissue or cells. It increases a lot the quantity of RNA you can extract. I usually work with as few as 15000 cells and I got enough RNA to measure OD, run in an gel and do RT. I would certainly add glycogen for a sample of 1 mg tissue.