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. 2017 Aug 10;48:41. doi: 10.1186/s13567-017-0445-2

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© The Author(s) 2017

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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.

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Detection of EGFP mRNAs of the recombinant HP-PRRSVs using northern blot analysis. Total RNAs from Marc-145 cells infected with six different recombinant HP-PRRSVs expressing EGFP and the parent strain were separated on a Tris–borate–EDTA–urea-15% polyacrylamide gel. The gel was transferred onto a piece of membrane (Hybond N+; Amersham). The blot was UV cross-linked using a cross-linking system (HL-200 HybriLinker; UVP), and DIG-labelled oligonucleotides were used as probes for EGFP sgRNA detection. The sequences for the probes used were as follows: SNB041-F, 5′-GTGAGCAAGGGCGAGGAG-3′; and SNB041-R, 5′-GTAGTGGTTGTCGGGCAGCA-3′. Numbers below the northern bands indicate relative levels of EGFP sgRNA of each recombinant virus compared to EGFP sgRNA of rHP-PRRSV/SD16/TRS2-EGFP. ImageJ software (NIH) was used to quantify the signal from the gel.