Hi Dr Sanjeevan,

Your question is not at all stupid, but very practical, at least in our context. I had a similar situation some years back and I decided to pause the RNA purification step after adding Isopropanol (was following TRI Reagent method for RNA prep) to it and carried it for the next 4-5 days without any coolant, during extreme summer conditions. Later, after reaching my lab I precipitated the RNA by high speed centrifugation, resuspended it in nuclease free water and stored aliquots in -80C. It worked absolutely fine. I suggest you the same.

Just to be sure, if you have some amount of any RNA sample to spare, then add 1 mL Isopropanol to it and leave it on your work bench (at RT) for couple of days. Later precipitate it, resuspend and check by gel or RT-PCR. You can check this with your Ethanol/ NaOAc also method . If any of the soultions work for you, send the samples in the same manner. Just make sure your plastic wares are Nucelase free, and do take care to seal the eppendorf cap with sufficient parafilm.

Let me know, if it works, 

best of luck,

SD

Dear Dr. Datta,

Thanks for the reply and happy to hear that it worked for you. Getting some level of confidence in me to send the samples in Ethanol/ NaOAc in RT. I was also thinking the same to do a small reaction leaving the RNA in tabletop for couple of days in Ethanol/ NaOAc. Let me see the outcome. Will surely let you know. 

Good Luck for your research

Saj

We have experience on keeping precipitated RNA at room temperature, it is not a problem, you can send it like you mention in na-acetate+ethanol, just make sure that your ethanol is 2.5 volumes of the sample and that your microtubes are well sealed (i.e. they do not leak)

best

Jose A.

It will be fine. Use screw-top microfuge tubes. Or better yet, send it as a lyophilized pellet. 

RNA later could also be alternative option if you are super concerned. That is of course if you do have a source for it.

Thanks Jose, Craig and Maha for the reply. 

Dear dr Sajeevan,

I'm currently facing the same issue, i actually need to ship RNA or cDNA samples at RT for sequencing, i would be grateful if you would please provide me more details about the stabilization protocol that you performed, and if it worked well with your samples what was the hold-over time recommended.

Thanking you in advance for your help

Best regards

Hi Imene,

Happy to help you. I isolated total RNA through the traditional method (refer to my publication for protocol) and checked the quality. Samples were shipped in normal Ice by adding 2.5 volumes of absolute ethanol. One of my friends carried it in luggage while flying and RNA was stable. The samples had around 7 RIN values after reprecipitating and got good RNA seq results. For the safer side, you can follow the kit method (Qiagen or Sigma) so that it will be more stable and can avoid RNase contamination by handing the traditional method. Keep in mind, if you handle the samples properly and perform a good RNA preparation, they will be stable. Even you can isolate and try to keep it in ice for 1 or 2 days and have a check before sending it. I don't know anything about your samples (plant or animal) also how the properties of the tissue. If you can share the information, I will be happy to help you more.

Good Luck!

Best,

Sajeevan

Thank you Dr Sajeevan for your prompt reply

I actually used the qiagen kit protocol for viral RNA isolation from animals swabs and tissues, if i convert my RNA samples to cDNA could i ship it at cold without adding ethanol? If so how long it wil hold-on? And for your samples did you add sodium acetate before shipping ? If yes at what concentration please?

Thanks again, and sincere appreciation