primer dissolved in wrong buffer (1M Tris HCl) - what to do now?? (Jun/18/2007 )
Hello everybody!
I was way too distracted and dissolved a lot of my primers in 1M Tris-HCl pH7.5 instead of 10mM Tris-HCl pH7.5.
And I dissolved them to a final concentration of 100pmol/µl. So the final 10x dilution would still leave me with 100mM Tris concentration...
Does anyone know what I can do??? Except ordering new primers? My PI will kill me...and right he would be. 
I also dissolved the plasmid DNA of a maxi prep in 1M Tris. Now I added 20 Volumes water and will try to precipitate. Will I get the DNA back??? Or is the HUGE amount of salt going to hinder the precipitation???
well you could try Ethanol percipitating the primers out. Wash it with 70% ethanol.
But the amount of primers are quite low. 200-250µg or so. Does it still work then?? With such small amounts of DNA?
Thermal DNA polymerases can tolerate a wide range of Tris concentrations.
Primers are too cheap that your PI would not bother you upon reordering.
The maxi DNA, can be precipitated and redissolved in the right buffer.
Don't worry, its all part of life.
But the amount of primers are quite low. 200-250µg or so. Does it still work then?? With such small amounts of DNA?
Hmm... on second though, don't bother. While possible, my idea isn't really workable. The primer concentration will probably be thrown off, and that will cause another headache.
As suggested by tfitzwater, you could try running the PCR with the primers as they are and see what happens
If things don't work, reorder said primers as Scolix has advised.
If you have to precipitate the primers, try using ammonium acetate, rather than sodium acetate, as it's much better for short pieces of DNA. 1/2 volume 7.5M ammonium Acetate, pH 7.5 to DNA, 2 volumes 95% EtOH. If the concentration is over ~50 ug/ml, precipitate at -70C for 15-30 min; if less, try overnight at -20C.
After I did something similar, I started putting my 1M Tris on the top shelf, so I'd really have to think about 
what I was doing.
Thank you all for your answers!! I called the guy from the sequencing facility yesterday and he told me, that the Tris concentrations shouldn't be a problem for the sequencing reaction, since the 100pmol/µl primer will be diluted 1:10 before sequencing and my maxi prep DNA is dissolved in water. So the Tris concentration in the final reaction mix shoulnd't ruin the whole thing.
But as you see: lots of "shouldn't" and no "will not".
And as to reordering --> 12 new oligos à 20-23 bases will be expensive. And my PI would be mad at me, because of my stupidity. 
Wee. I will let you know how the sequencing worked out. In case anyone is as scatterbrained as me and dissolves his/her primers in the wrong buffer!