Improved method for polynucleotide probe-based cell sorting, using DNA-coated microplates.

Abstract:

:We developed an improved method for cultivation-independent sorting of bacterial cells. The technique is based on labeling the target cells by in situ hybridization with polynucleotide transcript probes. Due to the probes' length, part of the probe remains outside the cell and can subsequently be used to capture the cells. Target cells are immobilized during a second hybridization step in microplates that are coated with DNA that is complementary to the probe sequence. The method was applied successfully to artificial mixtures of cells with polynucleotide probes targeting either rRNA, a plasmid-borne beta-lactamase gene, or a chromosome-borne glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Cells could be separated based on phylogenetic parameters (using rRNA-targeted probes) as well as on other DNA-encoded traits.

journal_name

Appl Environ Microbiol

authors

Zwirglmaier K,Ludwig W,Schleifer KH

doi

10.1128/aem.70.1.494-497.2004

keywords:

subject

Has Abstract

pub_date

2004-01-01 00:00:00

pages

494-7

issue

1

eissn

0099-2240

issn

1098-5336

journal_volume

70

pub_type

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