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100644 index 0000000000000000000000000000000000000000..ad81ff6bd64223b3afac5fa7507d29fc8d516915 --- /dev/null +++ b/assays/Maleckova2018_aba_mRNASeq/protocols/Illumina.md @@ -0,0 +1,3 @@ +# Preparation of Illumina libraries, and sequencing + +Libraries were prepared using the TruSeq Stranded mRNA Library Prep Kit (Illumina) following the Illumina TruSeq Stranded mRNA Sample Preparation Guide #15031047 Rev. E, with the following adaptations: 300 ng total RNA as the basis for library preparation, adapter index dilution to 50% with RNase-free water, and an additional purification step with AMPure XP (Beckman Coulter).The concentration of the resulting libraries was estimated using a NanoDrop 8000 (Thermo Fisher Scientific) and quality control was performed with a High Sensitivity NGS Fragment Analysis Kit (1 bp–6000 bp) DNF-474 (Advanced Analytical Technologies, Inc.). Based on quantification with a Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific), libraries were adjusted to 2 nM concentration before cluster generation with a HiSeq 3000/4000 SR Cluster Kit (Illumina). Twelve samples were pooled per lane and sequenced in 150 bp single-end mode on a HiSeq 3000 platform (Illumina). On average, over 32 million reads per library were obtained (Supplementary Table S1). \ No newline at end of file diff --git a/assays/Maleckova2018_aba_mRNASeq/protocols/RNAex.md b/assays/Maleckova2018_aba_mRNASeq/protocols/RNAex.md new file mode 100644 index 0000000000000000000000000000000000000000..8faf7f63f9bf7d557c078da0adb1029b8b8f0ee3 --- /dev/null +++ b/assays/Maleckova2018_aba_mRNASeq/protocols/RNAex.md @@ -0,0 +1,3 @@ +# Total RNA extraction + +Total RNA was extracted from ground leaf tissue using the Monarch®Total RNA Miniprep Kit (New England Biolabs Inc.) following the manufacturer’s instructions. Contaminating DNA was digested with DNase I (New England Biolabs Inc.). The integrity of input RNA was analysed on a 2100 Bioanalyzer (Agilent). \ No newline at end of file diff --git a/isa.investigation.xlsx b/isa.investigation.xlsx index c4c2b8c4f13d56351fa249134a567505587e1053..2dceaed5b38ad63ed9f2ee3f25ac5c1c4e81af6f 100644 Binary files a/isa.investigation.xlsx and b/isa.investigation.xlsx differ diff --git a/studies/Maleckova2018_aba/Maleckova_et_al-2019-Journal_of_Experimental_Botany-Transcript_and_Metabolite.pdf b/studies/Maleckova2018_aba/Maleckova_et_al-2019-Journal_of_Experimental_Botany-Transcript_and_Metabolite.pdf new file mode 100644 index 0000000000000000000000000000000000000000..b45e42bb067208768cd17d6b73030fd7438f03b3 Binary files /dev/null and b/studies/Maleckova2018_aba/Maleckova_et_al-2019-Journal_of_Experimental_Botany-Transcript_and_Metabolite.pdf differ diff --git a/studies/Maleckova2018_aba/README.md b/studies/Maleckova2018_aba/README.md new file mode 100644 index 0000000000000000000000000000000000000000..b074245fe34cf62f2c5f80395ca480d3515bbdb8 --- /dev/null +++ b/studies/Maleckova2018_aba/README.md @@ -0,0 +1,5 @@ +# Maleckova2018_aba + +Maleckova, E., Brilhaus, D., Wrobel, T.J., and Weber, A.P.M. (2019). Transcript and Metabolite Changes during the Early Phase of ABA-mediated Induction of CAM in Talinum triangulare. Journal of Experimental Botany 2: 16178–6596. + +https://doi.org/10.1093/jxb/erz189 \ No newline at end of file diff --git a/studies/Maleckova2018_aba/isa.study.xlsx b/studies/Maleckova2018_aba/isa.study.xlsx new file mode 100644 index 0000000000000000000000000000000000000000..fbe60363f9392b5eeb0d5842be36c6dae279ebe1 Binary files /dev/null and b/studies/Maleckova2018_aba/isa.study.xlsx differ diff --git a/studies/Maleckova2018_aba/protocols/.gitkeep b/studies/Maleckova2018_aba/protocols/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/studies/Maleckova2018_aba/protocols/plant_material.md b/studies/Maleckova2018_aba/protocols/plant_material.md new file mode 100644 index 0000000000000000000000000000000000000000..ddffa069e9552285422ad814c341e9227b4f7dc0 --- /dev/null +++ b/studies/Maleckova2018_aba/protocols/plant_material.md @@ -0,0 +1,3 @@ +# Plant material and growth conditions + +Talinum triangulare plants used in this study originated from two sub-sequent, controlled self-pollination events, increasing the homogeneity of the plant material. Seeds were germinated in multiplication substrate (Floraton 3, Floragard) and 4-week-old seedlings were transferred to pots with D 400 soil and Cocopor (Stender). Two weeks before the treatment, the plants were placed into a controlled-environment plant chamber (MobyLux GroBanks, CLF Plant Climatics) with the following growth conditions: 12 h light/12 h dark at 25 °C/23 °C. The light intensity at the leaf level was 150–200 μmol s–1 m–2. To avoid unintended induction of CAM by drought, all plants were watered as needed, with the same amount of water per pot. \ No newline at end of file diff --git a/studies/Maleckova2018_aba/protocols/treatment_harvest.md b/studies/Maleckova2018_aba/protocols/treatment_harvest.md new file mode 100644 index 0000000000000000000000000000000000000000..88610e2eaa87d032f8d4de1bddcd19f986e252f4 --- /dev/null +++ b/studies/Maleckova2018_aba/protocols/treatment_harvest.md @@ -0,0 +1,3 @@ +# Treatments and harvest of leaf material + +To determine a suitable ABA concentration, a range from 10 to 600 μM was tested (see Supplementary Fig. S1 at JXB online) prior to the work presented here. Using 2-month-old plants, two mature and fully illumin-ated leaves of each plant were treated either with (+/–) ABA [Sigma-Aldrich; 200 μM solution in 0.095% (v/v) methanol] or mock solution (0.095% (v/v) methanol). Both solutions contained 0.02% (v/v) Tween-20 (PanReac AppliChem). The first treatment took place after 4 hours in the light and the second treatment followed 4 hours later (Fig. 1). Treated leaves were harvested and snap-frozen in liquid nitrogen 40, 80, 160, 320, 640, and 1280 min after the first treatment. Two biological replicates originating from two independent plants were harvested per time point. The treated leaves from the same plant were pooled prior to all analyses. \ No newline at end of file diff --git a/studies/Maleckova2018_aba/resources/.gitkeep b/studies/Maleckova2018_aba/resources/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391