I don't know if I have RNA pellet or not - There are very little amount of RNA, Is there anything I can see to see the pell (Jun/05/2006 )
I’m extracting viral RNA from allantoidal liquid using trizol to do RT-PCR. There are always very little amount of RNA so I don’t see any pellet after precipitation, but most of the time I get a good band after PCR. The problem is that sometimes, I don’t have any band (my positive control is ok), if I repeat the RNA extraction (and RT-PCR), then I have the band. I’m almost sure that the problem is that sometimes I lose the RNA pellet when I wash with ethanol (75 %) or something like that, but as I never can see the pellet, I don’t know if I’m losing it or not. What can I do? Is there anything I can see to see the pellet??. Please help me!!!
after lysis and phase separation, you pipett the upper phase and add 1µl of a 100mg/ml glycogen as a carrier. Then isopropanol. So you'll see the pellet of glycogen, and by the way the RNA is inside.
Thank you very much... I’ll do it. Can I precipitate with ethanol or isopropanol after add glycogen? Is there any special treatment I have to do after that?
Again thank you very much!
you add glycogen and vortex few seconds. That ensure homogeneization.
After that point you add the alcohol of your choice and mix.
Then proceed as normal.