Greetings.
In my line of work DNA extraction is common, i had various experiences with both as buffer TE and water, the former is described as the better way to prevent DNA degradation but supposedly interferes with sequencing and PCR, nuclease free water theorically is safe too and doesn't has these disadvantages.
The experience i had in these cases with TE has been that it often doesn't reach renough concentration to interfere but the risk exist.
What are you opinions and experiences with the two resuspension options for DNA and which one of the two is generally recomended?
TE is better becausethe EDTA removes Mg and nucleases need Mg ions to be active and degrade dna. Water can become acidic and cause some depurination with time. The dilution of the dna in the pcr reaction and the fact that Mg is always in slight excess in a pcr reaction means that the edta will seldom have any effect
It mostly comes down to how long you want to store your samples. If you are instantly using them or at most freezing them overnight, using water is fine.
TE would only interfere with a PCR reaction if the volume of DNA added to the reaction is much higher than normal. But so long as the DNA added is low, then TE won't really interfere. And as everyone mentioned, your DNA will be more stable in TE.
- Badges
- Science method