Abstract:
:In the present study, EMA (ethidium monoazide) treatment was applied to a silty-sand reference soil prior to DNA extraction to enable a differentiation between dead and living cells. For this purpose, a reference soil was spiked with Listeria monocytogenes cells or cell equivalents, respectively. With the purpose of evaluating optimum treatment conditions, different EMA concentrations have been tested. However, the results remained largely inconclusive. Furthermore, varied dark incubation periods allowing EMA to penetrate dead cells did not allow the selective removal of DNA from membrane-compromised cells in downstream analyses. In contrast to undiluted soil, an effect of EMA treatment during DNA extraction could be observed when using a 1:10 dilution of the reference soil; however, the effect has not been sufficiently selective to act on heat-treated cells only. Although the application of EMA to soil requires further evaluation, the procedure harbors future potential for improving DNA-based approaches in microbial ecology studies.
journal_name
Curr Microbioljournal_title
Current microbiologyauthors
Wagner AO,Praeg N,Illmer Pdoi
10.1007/s00284-019-01772-ysubject
Has Abstractpub_date
2019-12-01 00:00:00pages
1425-1434issue
12eissn
0343-8651issn
1432-0991pii
10.1007/s00284-019-01772-yjournal_volume
76pub_type
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