I am trying to use a DSC to determine precise melting temperature of my DNA strands. I am using a TA instruments Q2000, and I am using Tzero pans/lids which I crimp together. Does it matter which pans I use? Should I be poking a whole in my lid? Liquid always seems to spill out but I am only using 2uL.
Note: melting temperature is the temperature at which 50% of DNA is dsDNA and 50% is ssDNA.
Hi
Use alumina crucible it wil be ceramic one and never stick.... Start from room temperature and maximum upto 200C with heating rate as for as minimum..... Try it now
Best
Arunprakash Vincent
Dear Hannah Kozl,
although the term "melting" would suggest DSC, usually the DNA melting determinations are carried out with UV/VIS and thermostatting. Please refer to https://www.mt.com/ch/en/home/library/guides/lab-analytical-instruments/uvvis_thermostating_guide.html, paragraph "3.4 Melting temperature of DNA".
Hope it helps,
kind regards, Samuele
Samuele Giani
I do not have access to a temperature controlled UV/Vis, which is why I am currently planning to use DSC. Also, do you have the direct pdf you can attach?
Thank you
I've never done a DSC for DNA, but I did it for proteins to measure at what temperature unfolding occurs. It might be similar to that.
We usually use hermetically sealed pans made of stainless steel. I don't think you should do a hole in lid unless you are heating your sample above water boiling point.
We use 50 microL of sample hermetically sealed and 50 microL of placebo solution in reference pan. We always use baseline correction.
Heating procedure: 30 C - 95 C with the heating rate 1 C/min.
Elizaveta Belova Have you ever done it without solution in the reference pan? That may be my problem; I was using a blank pan.
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