3 replies to this topic

[patty4150]

I have gotten some FAM/BHQ probes from a vendor and although they work initially, they go off quickly (within weeks.) The primers (from another source) seem fine.

I do not freeze thaw repeatedly, I protect from light, and I keep them stored at -80 long term. I am thinking the synthesis is not good. It has been a problem with all the probes we have ordered from this company.

Where do you get your real-time PCR probes, and have they been stable?

Edited by patty4150, 25 February 2010 - 01:12 PM.

[ivanbio]

Hi patty,

Assuming that your problem is the probes and not some other issue, a vendor I've used extensively for qPCR probes is Eurogentec. I've had probes last close to 2 years without any noticeable loss in signal. I never freeze and thaw more than once; I prepare a bunch of aliquots and store them frozen until I use them.

[pcrmossad]

patty4150, on Feb 25 2010, 10:12 PM, said:

I have gotten some FAM/BHQ probes from a vendor and although they work initially, they go off quickly (within weeks.) The primers (from another source) seem fine.

I do not freeze thaw repeatedly, I protect from light, and I keep them stored at -80 long term. I am thinking the synthesis is not good. It has been a problem with all the probes we have ordered from this company.

Where do you get your real-time PCR probes, and have they been stable?

I buy them from thermo electron. If you aliquote them and store them at -20 , theiy are fairly stable, and even worked after one year.

http://www.thermo.co...ome/1,,,00.html

[patty4150]

pcrmossad, on Feb 27 2010, 04:59 PM, said:

patty4150, on Feb 25 2010, 10:12 PM, said:

I have gotten some FAM/BHQ probes from a vendor and although they work initially, they go off quickly (within weeks.) The primers (from another source) seem fine.

I do not freeze thaw repeatedly, I protect from light, and I keep them stored at -80 long term. I am thinking the synthesis is not good. It has been a problem with all the probes we have ordered from this company.

Where do you get your real-time PCR probes, and have they been stable?

I buy them from thermo electron. If you aliquote them and store them at -20 , theiy are fairly stable, and even worked after one year.

http://www.thermo.co...ome/1,,,00.html

thank you both.  I am trying Biosearch next, and hoping for a better stability than seen with the previous manufacturer.