you should always dissolve pcr primers in TE. Water dissolves CO2 from the air to fprm carbonic acid which results in acid depurination of the primer and degradayion and the alkaline buffering of Tris prevents this. The EDTA chelates divalent ions  like magnesium so this prorects against nuclease degradation of the dna, There will be no effect on sequencing  which is a robust reaction and the small amount of primer added will have no effect on the larger amount of sequencing buffer with magnesium and manganese in it. 10mM TE is plenty to buffer the effect of CO2 absoeption

you should always dissolve pcr primers in TE. Water dissolves CO2 from the air to fprm carbonic acid which results in acid depurination of the primer and degradayion and the alkaline buffering of Tris prevents this. The EDTA chelates divalent ions  like magnesium so this prorects against nuclease degradation of the dna, There will be no effect on sequencing  which is a robust reaction and the small amount of primer added will have no effect on the larger amount of sequencing buffer with magnesium and manganese in it. 10mM TE is plenty to buffer the effect of CO2 absoeption

Agree with Paul, that primers are dissolved in TE, independent of their GC content.

The composition of 1X TE is 10 mM Tris-HCl (pH 8.0) containing 1 mM EDTA.

If cause a problem, you can send the product Gel  for sequencing, this is alternative way for synthesis the primer

If it is PCR primer MQ (DNAse RNAse free) water is fine as you will use the oligomer once to obtain your amplicon and the chances are that the primer will spend most of its life in -20 freezer.

As for the more expensive HPLC/Gel purified oligomers which are used in enzymatic reactions, primer extensions etc. TE is the best choice as it will maintain the stability of the oligomer for long period of time as Paul explained.

Finally, keep on mind that that there is many grades of chemicals you can purchase and it is also important to use them while they still within expiry date (TRIS and EDTA can expire too).

I agree with Stanislaw, to PCR you do not need TE buffer, in my lab we disolved PCR primers in MQ water and they last for long periods (years) at -20°C. I think that water is not your cause of degradation.

Cheers!

We always use nuclease free water to resuspend our primers. Primers are stable for years at -20C. 

We at Stanford (previous position) always resuspend in TE 10-1, this solves any complication with interference but still gives confidence of protection.

Hi, we resuspend our stock primers in TE 1X. Then, for working oligonucleotide solution we uses nuclease free water.

Tris EDTA buffer is the most suitable for dissolving primers as the buffer prevents degradation however there are chances of degradation with distilled water

For my stock primers I always resuspend it with TE, but for working primer I will use MQ water. Never have problem occur before for all my primers.

i have used RNase free water for primer dilution. you  can use distilled water for primer dilution.

The exact answer is given by Paul!

1XTE is the solution for your problem for 100%. You assumed it right but It wont interfere practically with your downstream applications.  

Keep your work area and bench/hood clean.

Water dissolves the oligos and you can store them in -20 for long.  But the temperature fluctuations and minimum # of thaw cycles can also damage the water dissolved oligos rapidly than that of TE dissolved. Even TE protects the oligos from enzymatic degradation if any. 

TE buffer is the best, it prevents degradation of primer during longer storage. If distilled water is used, must be endonuclease free. 

For long time storage, 1XTE buffer is the best. For routine work or short time use, nuclease free water can be used.

I always use nuclease-free water, but for DNA storage from patients I always use TE buffer. The question is how much time will you leave the primer unused? We have stock primers that have been prepared more than a decade ago that are diluted in nuclease-free water and still works perfectly. One problem that is worse than the type of solvent is thawing and refrezing many times the primers. Do you work with aliquots of your stock primer or dilute the stock to working conditions directly? 

Thank you all...

I generally use distilled water for my oligos dilution both stock and working concentration ( for the both application RT and conventional PCR). But my experience when i use nuclease free water for the GC rich oligos ( procured from bioserve and eurofins  ) life is 4-6 moths  and the other oligos were still stable (scince last 5 yrs).  

This time i will use TE and i share my experience..

Thank you all again..

i have always used nuclease free water( Thermos scientific)which has always worked fine for reconstituting my primers.However,making working solutions(aliquots)is vital to prevent contamination for good PCR results.Also,.It is important to store the nuclease free water vials in the refrigerator(at 4°c--20°c is just fine!).However,i will give TE a trial based on recommendations.

Keeping it simple, Tris resist pH change, EDTA chelates the metal ions....... so when it comes to dissolving oligonucleotides, u must always go for TE bufer.

I always use a nuclease-free water for preparation of primer solutions. I  did not come across any problem. You can use a nuclease-free water and avoid multiple freeze-thawing cycles for your primer solutions. You can prepare aliqoutes of primer solutions for single use.

It is difficult to aliqoute for single use when too small amount is required, for storing TE buffer is good. 

I generally prefer to use 1xTE buffer or Rnase containing water to dissolve and store DNA and plasmids. I prefer to use Diethyl pyrocarbonate (DEPC) treated water to dilute and store my primers. DEPC, also inactivates RNase enzymes in water and on laboratory utensils. You can also use autoclaved double dist water to store NA and primers. 

I've only just joined this discussion, but I am intrigued by the many mentions of freeze-thawing being able to degrade primers. Can anyone tell me how this happens, and how it has been demonstrated?

Generally for long term storage of primer use TE buffer and for working primer you can use nuclease free water. In TE buffer you can store your primer at -80 temperature for long time.

@Geoff

you are probably right about oligo degradation Geoff. IDT

https://eu.idtdna.com/pages/education/decoded/article/storing-oligos-7-things-you-should-know

thawed their oligos 30 times and found no difference in pcr efficiency although in pcr the primer is in huge excess so some damage would have gone unmeasured by this critereum.

taken from the above url

5. Freezing and thawing your oligos will have minimal impact on oligo stability.

It is not uncommon for some researchers to repeatedly freeze, then thaw their oligos while working in the lab. To address this, we functionally tested oligo stability after multiple freeze-­thaw cycles, using either nuclease-free water or IDTE Buffer as resuspension liquids (Figure 2). In our experiments, we found that 30 such cycles had no significant impact on oligo stability or function. This was true for both resuspension liquids tested (nuclease-free water and IDTE).

📷

Figure 2. Oligo stability is maintained after 30 freeze-thaw cycles. A standard scale IDT PrimeTime® qPCR Assay containing oligonucleotide primers and probes was hydrated in IDTE Buffer to 40X. The tube was frozen (–20°C) and thawed 30 times. At (A) 0 and (B) 30 freeze-thaw cycles, an aliquot of the assay was run against a validated universal human reference cDNA standard curve (0.005–50 ng) using TaqMan® Gene Expression Master Mix (Thermo Fisher). The oligo components of the assay were stable and functional for up to 30 freeze-thaw cycles, showing no impact on Cq value. (C) The PrimeTime qPCR Assay at 0.5 ng cDNA concentration is shown in a reference dye-normalized view (Rn).

Thank you Paul......