Using Math To Determine Purity In Unicorn ӒKTA Software

In this video I show how to add, subtract and divide curves to get additional insights to what going on in your purifications. You can determine if you have pure protein or nucleic acid using a 260/280 ratio for example and much more. DNA and RNA have an absrobance maximum at 260 nm while protein aromatic rings like tryptophan have an absorbance maximum at 280 nm Nucleic acids have absorbance maxima at 260 nm. If the absorbance ratio of 260/280 is 1.8 you can be more confident the peak is pure DNA, and 2.0 for RNA. The 260/280 ratio for protein is normally 0.6 and is informative but not a guarantee that it is pure. Proteins that have a low molar absorbance like BSA will have a higher 260/280 absorbance that looks like DNA.

Cytiva's Evaluation Module manual and curve arithmetic: https://www.oit.va.gov/Services/TRM/f...

More details about Nucleic acid quantitation: https://en.wikipedia.org/wiki/Nucleic...

For further reading generally on how to use the ӒKTA Avant see: https://cdn.cytivalifesciences.com/ap...

Please leave a comment below and good luck with your purifications!

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I don't work for Cytiva & I do not represent them in any way. The view's expressed here are my own.