Today these to PhD's told me that you lose up to 3/4 of RNA in a tissue sample by quick freezing it. I can't find any publications regarding those, but I'm not really proficient at looking for them. Is this true, and how can it be?
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#2
bob1
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Posted 14 May 2009 - 04:17 PM
I've never heard of that, snap freezing is a pretty common procedure for keeping samples for later extraction.
#3
sran
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Posted 14 May 2009 - 09:04 PM
If you snap freeze in liquid nitrogen and keep frozen, you should be fine.
Ahrenhase, on May 15 2009, 07:03 AM, said:
Today these to PhD's told me that you lose up to 3/4 of RNA in a tissue sample by quick freezing it. I can't find any publications regarding those, but I'm not really proficient at looking for them. Is this true, and how can it be?
#4
Vini
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Posted 15 May 2009 - 12:40 AM
sran, on May 14 2009, 10:04 PM, said:
If you snap freeze in liquid nitrogen and keep frozen, you should be fine.
Ahrenhase, on May 15 2009, 07:03 AM, said:
Today these to PhD's told me that you lose up to 3/4 of RNA in a tissue sample by quick freezing it. I can't find any publications regarding those, but I'm not really proficient at looking for them. Is this true, and how can it be?
many people store RNA after snap freezing. I cant imagine what could be the problem with that!!!
#5
nbhu
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Posted 15 May 2009 - 03:40 AM
I have actually done microarray analysis of the same tissue, one frozen, one snap frozen in nitrogen, and one in RNALAter and one in RNALater that was then frozen. Of these, RNA integrity got poorer as follows:
1. RNAlater and processed sstraight away (best RNA quality)
2. RNAlater and frozen/snap frozen
3. Snap frozen in nitrogen
4. Frozen tissue, no nitrogen.
REally it depends what you want to use the RNA from the tissue for, but if you wanted it for something as accurate as microarray analysis, i would reccomend some kind of RNA preservative like RNAlater (qiagen) though lots of companies have their own versions.
1. RNAlater and processed sstraight away (best RNA quality)
2. RNAlater and frozen/snap frozen
3. Snap frozen in nitrogen
4. Frozen tissue, no nitrogen.
REally it depends what you want to use the RNA from the tissue for, but if you wanted it for something as accurate as microarray analysis, i would reccomend some kind of RNA preservative like RNAlater (qiagen) though lots of companies have their own versions.
#6
Ahrenhase
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Posted 18 May 2009 - 03:47 AM
Ok, I was just wondering because I'm getting really low yields from ~30mg flash frozen tissue constructs (<100ng/ul). I ordered some RNAlater so it's good to hear people are having success with it. BTW, I'm using the RNA for qRT-PCR
