diff --git a/assays/ProteinAndStarchMeasurements/README.md b/assays/ProteinAndStarchMeasurements/README.md new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/ProteinAndStarchMeasurements/dataset/.gitkeep b/assays/ProteinAndStarchMeasurements/dataset/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/ProteinAndStarchMeasurements/isa.assay.xlsx b/assays/ProteinAndStarchMeasurements/isa.assay.xlsx new file mode 100644 index 0000000000000000000000000000000000000000..f4b3a8fba5380fac5773dab29986e9ef4e93792b Binary files /dev/null and b/assays/ProteinAndStarchMeasurements/isa.assay.xlsx differ diff --git a/assays/ProteinAndStarchMeasurements/protocols/.gitkeep b/assays/ProteinAndStarchMeasurements/protocols/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/ProteinAndStarchMeasurements/protocols/ProteinAndStarchMeasurementsProtocol.md b/assays/ProteinAndStarchMeasurements/protocols/ProteinAndStarchMeasurementsProtocol.md new file mode 100644 index 0000000000000000000000000000000000000000..5890e9eff0b1df0f67eb025a22930c0e538fe196 --- /dev/null +++ b/assays/ProteinAndStarchMeasurements/protocols/ProteinAndStarchMeasurementsProtocol.md @@ -0,0 +1,5 @@ +## Protein and Starch Measurements + +Approximately 25 mg of ground leaf material was used for extraction in 80% (v/v) ethanol with 10 mM 2-(N-morpholino)ethanesulfonic acid (pH 6). Extraction was performed three times with 500 µl extraction buffer while shaking for 1 hour at 90 °C. After each extraction step, samples were centrifuged at 12,000 g for 10 min and the supernatant was collected and pooled with previous fractions. From each sample, two extracts were prepared and stored at –20 °C until analysed. The protein concentration was measured using a Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). + +Pellets obtained during protein extraction were gelatinized with 300 µl 0.2 M NaOH at 90 °C for 40 minutes. After adjustment to pH 5–6, 200 µl hydrolysis buffer was added [20 mM acetate buffer, pH 4.8; 0.5 U α-amylase (10102814001, Roche), and 4 U amyloglucosidase (A7095, Roche)] and the mixture was incubated overnight at 37 °C. Glucose resulting from starch degradation was measured enzymatically: 10 µl of digest was added to 100 mM HEPES-NaOH (pH 7.5) with 10 mM MgCl2, 1 mM NADP+, 2 mM ATP, 0.1 U glucose-6-phosphate dehydrogenase (G6PDHII-RO, Roche), and 4 U hexokinase (11426362001, Roche). The final reaction volume was 200 µl and the increase of absorbance at 340 nm was monitored. \ No newline at end of file diff --git a/assays/TitratableAcidity/isa.assay.xlsx b/assays/TitratableAcidity/isa.assay.xlsx index 65d47771d540a71937a6455daf7140d8e6ba0eef..baccf6dea09270088bd0047c9c8cbbe41313807f 100644 Binary files a/assays/TitratableAcidity/isa.assay.xlsx and b/assays/TitratableAcidity/isa.assay.xlsx differ