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Pulsed-field gel electrophoresis (PFGE) analysis

BL Bojana Lukovic
IG Ina Gajic
ID Ivica Dimkic
DK Dusan Kekic
SZ Sanja Zornic
TP Tatjana Pozder
SR Svetlana Radisavljevic
NO Nataša Opavski
MK Milan Kojic
LR Lazar Ranin
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PFGE included 60 CRAB isolates randomly selected from all participating hospitals, with respect to the specimen types and gene content. PFGE was performed with a 2015 Pulsafor unit (LKB Instruments, Broma, Sweden), as previously described [28]. DNA restriction was done with ApaI enzyme (Thermo Scientific, Lithuania). The Lambda Ladder 48.5–727.5 kb PFG Marker (New England Biolabs, US) was used as DNA size marker. The stained gels were scanned using the Diversity Database software image-capturing system (Bio-Rad Laboratories Ltd., UK). The Dice coefficient was used to calculate similarities of the banding patterns with a tolerance of 1.5%, and the unweighted-pair group method using average linkages (UPGMA) was used for cluster analysis with BioNumerics software, version 4.0 (Applied Maths, St-Martens-Latem, Belgium). The isolates with more than 80% similarity in their DNA patterns were defined as genetically related and part of the same cluster.

Copyright and License information: The Author(s) ©2020
Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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