If properly stored in prescribed temperature, how long can a primer be used for PCR reactions?
Make some work solutions e.g. 100µl. So you will have no problems that your primers won't work!
No problem with storage, run a couple of PCRs and check concentrations. It should work.
I've had success with primers stored in TE at -20C for at least 5 years.
If they are stored in 10mM tris, 0.1mM EDTA at -20'C at a concentration of ~100uM, they should be stable for 5-10 years! The more dilute the primers are, the less efficient they tend to be after years of storage. A low concentration of EDTA can help prolong storage, but concentrations >0.5mM can inhibit PCR.
Mostly thawing, freezing, thawing....is the problem and then maybe the DNAses which can contaminate the primers by opening the tubes.
I've had success with primers dissolved in water and stored at -20C for at least 5 and half years years.
Hi,
we dissolve primers in MilliQ, autoclaved. make it up to 100uM as per sigma.
Make our desired aliquots for PCR into another tube (25uM) and store in -20C.
The main thing for stability is avoiding freeze thawing. Once u make your aliquots, leave your core stock frozen and it stays for more than 5 yrs!
Leave your aliquots aloso at 0C or -20C depending on usage. Its better to thaw them and use them on ice and hence they go on for a longer time than using it in room temp.
I thank all the members of this discussion for their valuable comments and sharing their experience. Researchgate rockz :-)
Bit late - but my personal record : approximately 15 years (100µM in TE-4 solution, stored at -20 and were defrosted several times)
first prepare a stock solution of higher concentration of the primer. this stock solution can be stored for indefinite time if properly freezed and stored at -20-40. For thawing, keep the vial in d.w.out of this stock make working aliquot for daily use which canbe stored separately.
Keep a high concentration (500uM) of the primers as stock and dilute to 50 or 100um and use for routine purpose. Avoid frequent freezing and thawing. Though the primers have long biological half-life they stil tend to degrade due to thawing
It can be used for several years, as suggested in above answers. I generally split the mother stock in smaller volumes and dilute these mini-stocks for use, as and when required.
Do already exist any papers in which the stability has been checked for more than 5 years at -20°C?
ThermoFisher has published a product stability study for qPCR assays. You can take a look. There are also several papers regarding the stability studies that other company did.
https://tools.thermofisher.com/content/sfs/manuals/cms_041915.pdf
Thank you Wilson Choo, do you have other studies in mind? You have already suggested how other companies published similar studies, I personally know that from IDT, nothing else? Perhaps something published in a peer reviewed journal?
Many thanks in advances
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