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Quantitative Reverse Transcriptase PCR

CK Courtney Kousser
CC Callum Clark
SS Sarah Sherrington
KV Kerstin Voelz
RH Rebecca A. Hall
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qRT-PCR was performed using an iTaq Universal SYBR Green One-Step Kit (Bio Rad) using 50 ng RNA with a total reaction volume of 20 μl. Protocol was followed according to manufacturer’s recommendations. FTR1 was amplified using the forward primer (5′-GTGGTGTCTCCTTGGGTGTT-3′) and reverse primer (5′-CCACCACGGTAGATGAGGA-3′). This was normalised to 18 s rRNA using the forward primer (5′-GGCGACGGTCCACTCGATTT-3′) and reverse primer (5′-TCACTACCTCCCCGTGTCGG-3′).

PvdS was amplified using the forward primer (5′-ACCGTACGATCCTGGTGAAG-3′) and reverse primer (5′- TGAACGACGAAGTGATCTGC-3′). PchA was amplified using the forward primer (5′- CTGCCTGTACTGGGAACAGC-3′) and reverse primer (5′-GCAGAGCAATTGCCAGTTTT-3′). These were normalised to rpoD using the forward primer (5′-GGGCGAAGAAGGAAATGGTC-3′) and the reverse primer (5′-CAGGTGGCGTAGGTGGAGAA-3′).

Copyright and License information: The Author(s) ©2019
Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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