RNA isolation / Glycogen - (Jul/01/2008 )

Hi,
I'm quite new in the field of RNA isolation and real-time PCR. I did some RNA isolation from animal tissue (Chmczynski & Sacchi method) and it did not cause to many troubles. Now I'm trying to isolate RNA from blood samples using Trizol. The results are quite good, when I isolate from 5-10 mln of cells. But I need to lower the cell number to 100 000 - 1 mln. And here problems occur. I read, that glycogen can faciliate the RNA yeald, but I don't know how much of it should I use and what concentration is the best. Has anyone got a clue how to use glycogen in this situation?
Thank You, in advance, for any help.

-ania_w-

QUOTE (ania_w @ Jul 1 2008, 11:40 PM)

Ambion sells Glycoblue, I always use that one with small amount RNA isolation. It is blue too, so helps my failing eyes

And check out these protocols:
http://search.vadlo.com/b/q?sn=158621799&a...cogen&rel=0
..

-cellcounter-

Thank You, for Your reply. It is really difficult to see the RNA pellet, so the GlycoBlue is a great idea. I've found ambion info about the proposed concentration used in experiments. But I was more curious how much of it people really use in labs. Could You tell me how much of it You use? I would like to ask about drying RNA. Sometimes it is hard to get rid of the ethanol completely are there any good and safe ways to do so?

-ania_w-

QUOTE (ania_w @ Jul 7 2008, 12:23 AM)

1. They suggest 1 ul per sample. I go with the same. Sometimes if I feel insecure, I add two

2. getting rid of ethanol is a trick you develop! Several ways, not going to go in detail at this point.

-cellcounter-

Thanks once again, Cellcounter. I'm going to follow Your advice and hope it will work for me too

-ania_w-