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RNA isolation from locus ceruleus - (Jul/06/2006 )

I am a grad student and new to RNA isolation techniques. I have to extract total RNA from LC isolated from mice. LC isolation protocol that we follow takes a minimum of six hours.I dont want to culture the LC after I isolate them . I want to extract RNA directly from the cells immediately after isolation. My concern is that will I get undegraded total RNA from the isolated LC after the long LC isolation procedure.

-Seema Haridas-

QUOTE (Seema Haridas @ Jul 6 2006, 12:30 PM)
I am a grad student and new to RNA isolation techniques. I have to extract total RNA from LC isolated from mice. LC isolation protocol that we follow takes a minimum of six hours.I dont want to culture the LC after I isolate them . I want to extract RNA directly from the cells immediately after isolation. My concern is that will I get undegraded total RNA from the isolated LC after the long LC isolation procedure.


What is LC?

-Minnie Mouse-

LC is the locus coeruleus...

for extraction : i don't know the procedur of isolation.
But try towork on clean area willdecrease the risks of degraded RNA.
Before extraction, rinse well the sample as the surface is always damaged and slight degraded RNA is present.
Then homogenize on ice or in trizol with precautions.
Normally the rinse procedure decreases degraded RNA presence.
If you can, do the extraction at lowest temperature possible (in a cold chamber for whole extraction is highly recommended).
you may also use RNA later if applicable (i don't use it but know it's a preservative for RNA integrity)

-fred_33-