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2 step RNA isolation


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#1 Rutger

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Posted 15 June 2009 - 12:21 AM

Hello,

i am doing experiments for which i have stimulated cells after a 1 and 2 week culture (human cells)
After this period i want to isolate RNA from them to do a qPCR following RT-PCR.
Problem is, as a student i cant always get space in the Molecular lab, and my experiment ends on
different time intervals. If possible i would like to homogenize my cells first and do the isolation later
with all my samples together, so i only need to use the molecular lab once for a long time.

I am using Aurum Total RNA Mini Kit, spin protocol for cultured cells, bateria and yeast.


Is it possible to just homogenize the cells and then freeze them in lysis solution ( with 1% B-mercapto-ethanol)
Or is freezing the cells just after harvesting a better idea?

Thx in forward,

Rutger

Edited by Rutger, 15 June 2009 - 01:01 AM.


#2 molgen

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Posted 15 June 2009 - 01:40 AM

First, I'd ask bio-rad tec support if you can freeze the cells in lysis solution.

Second, I usually freeze the cells just after harvesting and never had a problem with downstream applications.

#3 Rutger

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Posted 15 June 2009 - 01:50 AM

First, I'd ask bio-rad tec support if you can freeze the cells in lysis solution.

Second, I usually freeze the cells just after harvesting and never had a problem with downstream applications.


thx, i think i will just freeze cell pellet then and use that for RNA isolation.




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