RNA Storage : Short and Long-term Storage

[bioinfofreak]

RNA Storage

*RNA may be stored in several ways.
Short Term Storage

*Store RNA in RNase-free H2O (with 0.1 mM EDTA) or TE buffer (10 mM Tris, 1mM EDTA).

*RNA is stable at -80° C for up to a year without degradation.

*Note that Magnesium and other metals can catalyze non-specific cleavage of RNA. These can be chelated by the addition of EDTA if RNA is to be stored and retrieved intact.

* Important!: use an EDTA solution known to be RNase-free for this purpose (older EDTA solutions may have microbial growth which could contaminate the RNA sample with nucleases).

* RNA solubilized in formamide solution can be stored at -20°C without degradation for at least 1 year (Chomczynski, 1992).


Long Term Storage of RNA

RNA samples may be stored at -20°C as ethanol precipitates for long periods of time.

However, accessing these on a routine basis can be a nuisance, as the precipitates must be pelleted and dissolved in an aqueous buffer or water (Rnase-free) before pipetting.

[moleculardude]

Thank you for a helpful protocol!

[khalsakh]

hi

i worked on RNA ONLY, for 3.5 years.... it was very hectic time but at some how v interesting.. if u need any help that i can offer i am welling to do,, and every body is wellcome....

[admin]

Hello Khalsakh!

that is great news! we really need more RNA people. Any recommendations on storing RNA?

[khalsakh]

hi ...

I used to store the cell line pellete in ultraspec after isolating the wbcs by ficoll. Then store it at -80. whenever i need the RNA i extract some aliquote from the pellete. I was using the samples for a research, therefore i need sometimes later to go back and extract some more. This procedure takes longe time BUT keeps us in a very safe side not to loose the samples after a while due to the RNAase activity. I am still keeping the pelletes till now( 5 years since taking the samples)....

[moleculardude]

Great information Khalsakh!
Wow 5 years of RNA storage is amazing!!

thanks for sharing. Do you use formaldehyde to store DNA or not?

thanks

[tomoe]

hi....

how many time we can freeze and thaw RNA?

thank you

[neeta]

hi,,


I m isolating total RNA from bacterial cell(ecoli) using trizol method referred by life science technologies bt i m not getting a good quality RNA..The absorbance of the RNA sample after quantitating comes very poor...please tell me some other method if u know for isolating RNA from the RNA sample.....

thankyou

[khalsakh]

Trizol is very widely used and recommended by many researchers. I never used it so far. Did u managed to Free your working area and materials from the RNase? ur gloves and glasswares? ur working solutions?

There are also some inhibitors and interferences in the reagents of the extraction; organic artifacts. U might need to take precautions at that stage; e.g ethanol...etc.

[Aurora Borealis]

We isolate RNA from leucocytes. I have no experience with other types of cells, but with leucocytes we have no problems getting fine quality and quantity of RNA. We use RNeasy Mini Kit from Qiagen - a column based system. It's easy and quick.

We simply store the RNA at -70oC in the water it's eluated in. We have RNA from years and years back, and it's still fine. I believe that if the RNA is isolated with clean equpiment, in clean working area and with good routines it can be stored a long time, unless you are thawing and freezing it over and over again.