dNTP degradation - (Jan/31/2006 )
Hi,
I just started honors and I was just woundering how dNTPs actually degrade if they are not kept at -20 degrees or after a number of freeze thaw cycles.
Thanks!
They are usually quite hardy... freeze thaw does not degrade them - as long as its not tooo many times. But they are best strored at -20 degress.
aliquot them and store at -20° and you'll be able do to lots of bright PCR.
considering ligation buffer, it's said that the ATP needed for efficient ligation is degraded by multiples freeze thaw cycles. So for dNTP i consider that point too.
But 10 freeze thaw cycles would not stronlgy degrade dNTP... I mean that you can still aliquot them.
oh, ok,
so what is actually the main reason that people aliquot it (i guess to reduce contamination and just for the convenience) and store it at -20?
considering ligation buffer, it's said that the ATP needed for efficient ligation is degraded by multiples freeze thaw cycles. So for dNTP i consider that point too.
But 10 freeze thaw cycles would not stronlgy degrade dNTP... I mean that you can still aliquot them.
when i said it's still time to aliquot, i did not mean (sorry for misunderstanding if so) that dNTP were 100%ok in your tube. I meant that if aliquoted, you will preserve remaining aliquots for future applications, which would not be if you don't aliquot.
Three reasons for aliquoting : preserve the dNTP, avoid contaminations of a full vial, and it's more convenient to thaw 10µl than 500µl...