add fawke study and qpcr protocol

studies/Fawke-2019/protocols/qRT-PCR.md

+## Quantitative reverse transcription polymerase chain reaction (qRT-PCR)
+
+Total RNA was extracted from plant material using Qiagen RNeasy Plant Mini Kit (Qiagen), including 1% (v/v) β-mercaptoethanol in the extraction buffer. RNA was then reverse-transcribed to cDNA using the Roche Transcriptor First Strand cDNA Synthesis Kit. qPCR was performed in 384 well plates using a Roche LightCycler 480 SYBR Green I Master Mix in a Roche LightCycler 480 II machine. Three technical replicates were performed for each sample. Normalization of crossing point-PCR-cycle (Cp) values to an internal control was performed against NbEF1a, NbF-BOX or NbL23 (Liu et al., 2012) for quantification of *N. benthamiana* transcripts and against PiWS21 (Yan & Liou, 2006) for *P. infestans* transcripts.
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