For how much time cDNA can be stored at -40 deg. Is it possible to store cDNA for nearly 6 months or above? Thanks for advise in advance
I am not saying about DNA my question is cDNA ie After Reverse transcription of RNA for how long cDNA can be stored
Stability of cDNA is more compare to RNA. You can store it in -20 C for more than 1 year. Even in 4 C its stable upto 6 month. For PCR & qRT PCR its ok.
From my experience cDNA appears to be much more stable than RNA. It can be stored at -20C for quite long time. I have used cDNA after 2,5year storage (-20C) and it worked just fine. To be extra careful you can store your cDNA in TE buffer.
From my experience, concentrate cDNA can be store at -20°C for a long time (more than 6 month, sure). Usually, what I do for sensitive experiments, such as qPCR, I dilute cDNA extemporaneously, and I don't keep it for another experiment. Diluted cDNA does not like -20°C storage and too many freeze-thaw cycle.
cDNA is more stable as compare to RNA. you can freez it at -20 for a long time, more than 5 years.
cDNA can be store at -20°C for a long time. I suggest you to avoid repeated freeze thaw. Its better to store as aliquots.
Agreed with Jaya Gosala. While cDNA can be stored in -20 for a long time, repeated freeze-thaw will cause it to degrade. If you want to use it to run PCR, you should store them in aliquots.
Sure! -20ºC is okey, and -40ºC too (maybe better). DNA can be degraded by DNAses and also by acid hydrolysis. You can avoid the first issue mainly working with DNAse free materials and reagents. To avoid the second issue, aliquot your DNA to thaw-freeze your DNA as little as possible. You'll also reduce the spontaneous acid hydrolysis of your DNA resuspending it into 10 mM Trsi-HCl pH 8.0 instead of water.
I have DNA preps that have been stored at -20ºC for over 20 years, so it should not be a problem. But I agree it would be wiser to not use a frost-free unit that cycles.
Aliquot, freeze (-20 C is good enough), forget...and use as required. DNA will stay loyal.
Dear B.A. Padder
It is better to aliquot into 2-4 microtupe, so after that, you can store at -20--80 oC for over 5-10 years (sometimes for over 20 years).
All the above are correct. Also, avoid divalent cations. Good clean preps last longer. Desalting on a spun column for final buffer exchange into 10 mM Tris. pH 8 would be nice.
In my hands after cDNA prep using Qiagen kit I stored cDNA as such in -20 which lasted a very long time, at least 3 years without degradation. Just have to take care of using very clean tubes, tips and buffers during cDNA prep.
I cryopreserved whole, functional lymphocytes and thawed them fifteen years later for purposes of a (successful) long-term study that showed no significant losses in viability or function. Proper freezing, storage, and thawing is key.
It is fine at -40 C, but store in aliquots and avoid repeated freeze-thaw.
From my experience cDNA can be stored at -20°C for more than a year, and can be used in sensitive experiments like q-PCR. If you need to dilute your cDNA, I suggest you to use just a small volume at time. Good luck!!
Yes, you can store cDNA in -40 for more than 6 months even several years. You can also store in -80 for very long period. cDNA quality will remain good as long as you handle it well, preferably make sure to thaw on ice.
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