I am currently working on the epigenetic (methylation) aspect of coronary artery disease. After short-listing a list of CpG sites with the help of microarray data I am currently working on HRM to semi-quantitate the extent of methylation of few sites.
The standards procured are mixed in equimolar concentration and 0% ,25%, 50%, 75% and 100%. All the input DNA are maintained at 20ng/ul.
However, in the process of standardization, I am facing a concurrent issue where a few samples are consistently coming as beyond 100%. How can I interpret this data?
Shyamashree Banerjee
Can you elaborate what you mean by this - "The standards procured are mixed in equimolar concentration and 0% ,25%, 50%, 75% and 100%".
Additionally, did you use nanodrop for DNA quantification?
Hello Abhijeet Singh ,
Yes I did use nanodrop for quantification
Also, when I mean mixed in equimolar concentration, I mean if my 0% quantifies 20ng/ul and 100% is 20ng/ul, for 50 % I will take 1:1 ration (10ng of 0% and 10ng of 100) for 25% (5ng of 0% and 15ng of 20ng of 100%)
Hope this clarifies
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