diff --git a/assays/RNA_extraction/protocols/trizol_RNA_extraction.txt b/assays/RNA_extraction/protocols/trizol_RNA_extraction.txt index d80fb95dc1c55354483ac232bd4fa66d923f3c29..160b4a9ab47dce00c4a30b521ec66e050a2b9c52 100644 --- a/assays/RNA_extraction/protocols/trizol_RNA_extraction.txt +++ b/assays/RNA_extraction/protocols/trizol_RNA_extraction.txt @@ -1,9 +1,5 @@ TRIzol RNA extraction -- Place 2 3mm metal beads into 2.0 ml tube -- Collect samples directly into the tubes on liquid nitrogen -- Store samples at -80°C until needed - - Grind your samples 2 x 45sec at maximum frequency (30 Hertz) in TissueLyser (QIAGEN), use pre-cooled adapters - Add 500 µL of TRIzol to the frozen samples, close tubes and mix vigorously until the sample is homogeneous - Incubate the samples for 5 min at room temperature diff --git a/studies/gene_expression_eam7_plants/protocols/plant_material_growth_conditions.md b/studies/gene_expression_eam7_plants/protocols/plant_material_growth_conditions.md new file mode 100644 index 0000000000000000000000000000000000000000..88974d69bfb8f2cdf35b05b4c0edee43fee5ed68 --- /dev/null +++ b/studies/gene_expression_eam7_plants/protocols/plant_material_growth_conditions.md @@ -0,0 +1,35 @@ +**Plant material and growth conditions** + +**plant material* + +species: Hordeum vulgare +genotypes: + +- BW: Bowman, spring cultivar +- BW(Ppd-H1): Ppd-H1 introgression line BW281 in BW background, Druka et al. (2011) +- BW(eam7): eam7 introgression line BW281 in BW background, Druka et al. (2011) +- BW(Ppd-H1, eam7): cross of the two introgression lines + + +**growth condtions** + +- short day (SD): 8h light / 16h dark, 20/16 °C, 60% humidity, ~ 250 PAR) +- QuickPot 96T trays (HerkuPlast Kubern GmbH, pot volume 75 cm^3) +- Einheitserde ED73 (Einheitserde Werkverband e.V.) with 7% sand and 4 g/L Osmocote Exact Hi.End 3-4M, 4th generation (ICL Group Ltd.) + + +**sampling** + +sample every 2 hours over 24 hours: + ZT0 (lights are turned on) + ZT2 + ZT4 + ZT6 + ZT8 (lights are turned off) + ZT10 + ZT12 + ZT14 + ZT16 + ZT18 + ZT20 + ZT22 \ No newline at end of file