DNA fragmentation in a steady shear flow.

Biomicrofluidics

Department of Chemical Engineering and Materials Science, University of Minnesota, Twin Cities, 421 Washington Ave SE, Minneapolis, Minnesota 55455, USA.

Published: September 2022

We have determined the susceptibility of T4 DNA (166 kilobase pairs, kbp) to fragmentation under steady shear in a cone-and-plate rheometer. After shearing for at least 30 min at a shear rate of , corresponding to a Reynolds number of and a Weissenberg number of , % of the sample is broken into a polydisperse mixture with a number-averaged molecular weight of  kbp and a polydispersity index of , as measured by pulsed-field gel electrophoresis (with a 95% confidence interval). The molecular weight distributions observed here from a shear flow are similar to those produced by a (dominantly extensional) sink flow of DNA and are qualitatively different than the midpoint scission observed in simple extensional flow. Given the inability of shear flow to produce a sharp coil-stretch transition, the data presented here support a model where polymers can be fragmented in flow without complete extension. These results further indicate that DNA fragmentation by shear is unlikely to be a significant issue in microfluidic devices, and anomalous molecular weight observations in experiments are due to DNA processing prior to observation in the device.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9616606PMC
http://dx.doi.org/10.1063/5.0109361DOI Listing

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