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zc=ccP`Zw%{#y<=DZ;j=DhyAC_|B3#y#{5Pbko-;Q&x-RW{LiHL8-7UoYpMKv2|!*7 U{;$1;`EUT~d03I|<ln{rAJy81v;Y7A literal 0 HcmV?d00001 diff --git a/studies/Fawke-2019/protocols/.gitkeep b/studies/Fawke-2019/protocols/.gitkeep new file mode 100644 index 0000000..e69de29 diff --git a/studies/Fawke-2019/protocols/qRT-PCR.md b/studies/Fawke-2019/protocols/qRT-PCR.md new file mode 100644 index 0000000..9ca8775 --- /dev/null +++ b/studies/Fawke-2019/protocols/qRT-PCR.md @@ -0,0 +1,3 @@ +## 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. \ No newline at end of file diff --git a/studies/Fawke-2019/resources/.gitkeep b/studies/Fawke-2019/resources/.gitkeep new file mode 100644 index 0000000..e69de29 -- GitLab