[RASMB] AUC measurements in CsCl?

[John Sumida]

Dear RASMB,

 

First I have to thank all those who curate this list server as over the last
10 years as a manager of the lab I run, it has and continues to prove a
hugely valuable resource for me and the people I serve.

 

My question today has to do with the analysis of DNA and it stems from a
user interest in performing an AUC equilibrium experiment in the presence of
CsCl.  My experience to date has been with nanomaterials and proteins so DNA
is a new material for me, however I am under the impression that running AUC
samples in presence of CsCl is problematical because of the gradient CsCl
itself will generate.  

 

The estimated mass of the species of interest is in the range of 10 mega
Daltons; this is also a size regime that is new to me.

 

The objective of the experiment is to characterize nucleic acids extracted
from cells which will have a label increasing the mass of the nucleic acid
depending on the number of times it has been replicated.

 

Two questions:

Is CsCl necessary to determine the size distribution of molecular species
>10MDa?

Can a CsCl experiment be performed using equilibrium measurements?

 

Thank you for considering my question.

 

Best regards,

 

John Sumida

Molecular Analysis Facility

University of Washington

 

In many spheres of human endeavor, from science to business to education to
economic policy, good decisions depend on good measurement.

Ben Bernanke



 

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