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Effect of rotor type on the separation of isotope-labeled and unlabeled Escherichia coli RNA by isopycnic density ultracentrifugation

Author:
Herrmann, Elena, Koch, Patrick, Riedel, Christian U., Young, Wayne, Egert, Markus
Source:
Canadian journal of microbiology 2016 v.63 no.1 pp. 83-87
ISSN:
1480-3275
Subject:
Escherichia coli, RNA, density gradient centrifugation, isotope labeling, microbial communities, stable isotopes, urea
Abstract:
Separation of differentially isotope-labeled bacterial RNA by isopycnic density gradient centrifugation is a critical step in RNA-based stable isotope probing analyses, which help to link the structure and function of complex microbial communities. Using isotope-labeled Escherichia coli RNA, we showed that an 8 mL near-vertical rotor performed better than a 2 mL fixed-angle rotor, thereby corroborating current recommendations. Neither increased concentrations of formamide nor urea in the medium improved the separation results using the fixed-angle rotor.
Agid:
5601800