By accident I left my cell material after using TRIzol at room temperature for about 20 hours. When I realized it was left outside, I immediately put it at -70 C. However, I think my cell material might not be usable anymore.
At 4°C for sure you could be safe. In my opinion at the present is convenient to proceed with the extraction. Storage at -70° now is unnecessary, because the material could be degraded.
I have had my cells in trizol for over 4 weeks at -20 degs and had no problems with further expts. as serena says -70 is not required
I'm not sure about cells but I have extracted honeybee parts, including guts and brains into trizol and due to the numbers this can take 8-12 hours, after freezing them I sometimes had to thaw them and continue extraction and they have been been fine.
I have had tissue in trizol during two days at room temperature and the RNA extraction was ok. This is not the best way but in general the material doesn´t look higly degrated. The RT were enough useful to clone the genes that I had expected to obtain from this samples.
U shud proceed immediately. Dont even keep at -70 now. But now the chances of good quality RNA are very less. All the best !!!!
I have kept pituitary extracts in TRIZOL for few hours at room temperature but it is not advisable since RNA quality and TRIZOL both degrades gradually with time. However you should check yield of mRNA of your interest...........if they are present in bulks in fresh extract.......chance of getting a working amount is still there but don't store these samples further............even storing at lower temperatures had disadvantages......avoid them. Best of luck!
It's not a problem if you left Trizol like this.But the efficiency may decrease.It's not advisable to left like this.
Hi Katrina,
I have tested the RNA degradation in room temperature Trizol versus -70C Trizol. I performed the extraction after intentionally leaving mock Trizol samples at room temperature and frozen at -70C for a week each. I ran the samples on agarose gel and I could see no difference whatsoever. The Trizol very efficiently inhibits RNase enzymes so there is little to no visible sample degradation after an entire week. From my test, I assume there is little spontaneous RNA hydrolysis in the aqueous phase of Trizol. From my GAPDH reading on qRT-PCR on my two samples, the GAPDH mRNA levels were not affected. This is not proof but this will reassure you a bit.
Jean-Charles
Hello Katrina,
If the biological sample is efficiently lysed in TRIzol and the reagent can inactivate the nucleases, RNA can be safely stored for 3 or 4 days at room temperature (20-25ºC).
http://tools.lifetechnologies.com/Content/SFS/ProductNotes/F_071215_Trizol%20and%20Trizol%20LS.pdf
Also you can preserve the RNA integrity using the RNAlater reagent from Qiagen, which immediately stabilizes RNA in tissue samples and provides a safe handling at room temperature for a longer time.
I have a question,
In order to preserve RNA with protocol above, is chloroform needed? (I don't mean extraction)
As well, which one is better? Trizol or RNA later?
Thanks
What if there is a gel that store your sample in room temperature forever and allow you to do all omics analysis
What if the cells in trizol is kept in -20 for a week and then transferred in -70? How long can we keep cell in trizol in -70?
For how many days i can store plant tissue sample in trizole at 4 degree Celsius?
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