Capillary Zone Electrophoresis with Fraction Collection for Separation, Culturing, and Identification of Bacteria from an Environmental Microbiome

Analytical Chemistry
Bonnie Jaskowski HugeNorman J Dovichi

Abstract

Capillary zone electrophoresis (CZE) can produce high-resolution separations of biological samples, including microbial mixtures. The study of complex populations of microorganisms using CZE is limited because most detectors have limited sensitivity, are destructive, and provide limited information for microbial identification. To address these issues, we developed an integrated capillary zone electrophoresis apparatus to fractionate bacteria from complex mixtures. We deposited fractions onto nutrient agar in a Petri dish for microbial culturing, and we subjected the observed colonies to Sanger sequencing of a phylogenetic marker, the 16S rRNA gene, for microbial identification. We separated and cultured both a single bacteria species, the model Gram-negative organism Escherichia coli, and a complex environmental isolate of primary sewage effluent. Sequence analysis of the 16S rRNA genes from this mixture identified 15 ± 5 distinct bacterial species per run. This approach requires minimal manipulation of microbial populations and combines electrophoretic fractionation of bacterial cells with automated collection for accurate identification of species. This approach should be applicable to microorganisms in general and may enabl...Continue Reading

References

Jan 1, 1991·Journal of Bacteriology·W G WeisburgD J Lane
May 1, 1990·Journal of Virological Methods·J L Sloyer, M E Bayer
Nov 1, 1993·Bio/technology·R C Ebersole, R M McCormick
Jan 7, 2000·Analytical Chemistry·D W ArmstrongD J Westenberg
Mar 4, 2000·Analytical Chemistry·S N KrylovN J Dovichi
Jul 1, 1953·Applied Microbiology·R S HARTMANR D RODGERS
Sep 22, 2009·Electrophoresis·Esra Acar Soykut, Ismail Hakki Boyaci
Dec 5, 2009·Electrophoresis·Michael W VannattaNorman J Dovichi
Apr 13, 2010·Nature Methods·J Gregory CaporasoRob Knight
Jun 5, 2012·Journal of Bacteriology·Eric J Stewart
Nov 30, 2012·Nucleic Acids Research·Christian QuastFrank Oliver Glöckner
Dec 3, 2013·Nucleic Acids Research·Pelin YilmazFrank Oliver Glöckner
Dec 18, 2013·Electrophoresis·Ewelina Dziubakiewicz, Bogusław Buszewski
Aug 28, 2014·Talanta·Bonnie Jaskowski HugeNorman J Dovichi
Jul 7, 2015·Nature Reviews. Microbiology·Martin Ackermann
Mar 5, 2016·World Journal of Microbiology & Biotechnology·Agnieszka Cydzik-Kwiatkowska, Magdalena Zielińska

❮ Previous
Next ❯

Citations

Mar 24, 2021·Analytical and Bioanalytical Chemistry·Ziting Gao, Wenwan Zhong

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.