real time pcr with RNA co-precipitated with glycogen - (Oct/07/2007 )
i want to know if rna co-precipitated with glycogen by trizol reagent will have any problem in real time pcr
thanks in advance
Hi yasd, I use Tri reagent for RNA isolation and usually precipitate it with isopropanol. Also, I have sometimes isolated DNA by precipitating with sodium acetate and glycogen, using isopropanol in the next step. In both cases I have used my samples for quantitative PCR with no problems. However I would recommend you to follow Trizol protocol without changes, since precipitation works fine and the final yield is good.
thanks Veronica for your suggestion, do you have any idea for how long tri reagent is stable after opening a new lot....our lot is nearly 5 months old and when i checked its ph it was around 3.5 , i think it should be 4.5
can u help me in this regard please...
I used glycogen to co-precipitate DNA from a very small amount of cells in Trizol. I used in real-time PCR and no problem. I don't think it would make difference with RNA.
Anyway for small amounts of RNA we use Trizol and RNA-carrier based on a RNA homopolymer from a Czech company (but they don't have web pages in english as far as I know), which is suitable for subsequent PCRs.