Half life of PCR reagents at different temperatures

[Guido D. Núñez-Mujica]

Greetings.

I am interested in finding out if somebody knows about a study or has practical experience about the half life of PCR reagents at different temperatures. I am thinking about cheap PCR applications for undeveloped countries, and one of the drawbacks in extreme environments can be the stability of the reagents. I am not really concerned about Taq, coming from an extremophile, but the ATP is extremely labile. I am not sure about the stability of nucleotides neither.

I think that if it would be possible to reduce refrigeration costs keeping at low temperatures only what is necessary the costs of freezing the material could decrease and therefore, would be cheaper for people in rural environments.

Thanks in advance

[Mario Raya]

... I am not really concerned about Taq, coming from an extremophile,...

But you should be concerned about the TAQ, if not stored properly its half-life is hours and, considering costs, is the most expensive reagent. The dNTPs are also sensitive, but not as the TAQ. The other reagents are usually buffers and salts, you don't have to worry about those.

At BIO2007, i saw a company (i don't remember the name) that was selling water with some other reagents, they claimed that those would keep the reagents for PCR stable at room temperature and give a longer half life at 4C, sadly we were there as the booths were closing, but i think i still have some info at work, that may be a good option.

Mario

[Tom Knight]

Check out GE HealthCare / Amersham puRe Taq "Ready-to-go" PCR beads,
which are dried down room temperature stable PCR enzymes, dNTPs,
buffers, etc. Just the thing for PCR on the go. Product 27-9559-01.
There's a patent on making these, but the key ingredient is trehalose,
which stabilizes protein mixtures in the dry state.

[Mario Raya]

(i apologize for the self-promotion)

I work at MassBay Community College, we are organizing our first conference about forensic DNA, it will be on November 7th and 8th, please, see the attached image for more information.

Some information about Daniel Vanek.... http://www.praguepost.com/articles/2007/05/23/piecing-together-the-dna-puzzle.php

A recent article about our forensic DNA program... http://www.boston.com/lifestyle/family/articles/2008/09/23/revealing_hidden_roots/

[Guido D. Núñez-Mujica]

Thanks a lot for your advice, it really will be helpful and will give us ideas.

[Guido D. Núñez-Mujica]

Great advice, Mario. I was totally clueless about it. It is great to have the experience and help from the whole group, it will make things easier.