Arabidopsis seeds were surface-sterilized in 50% bleach with 0.01% Triton X-100 for 15 min and washed five times with sterile, doubly distilled H2O before plating on MS medium (4.3 g MS salt, 10 g sucrose, pH 5.7, 8 g phyto agar per liter). After 3-4 days of cold (4 °C) treatment, the plates were wrapped in foil and kept in at 24 °C in an incubator before the phenotypes of seedlings were analyzed. For propagation, seedlings were transferred from plates to soil (Pro-mix-HP) and grown to maturity at 22 °C under 16-h light/8-h dark cycles. Ethylene treatment of Arabidopsis seedlings was performed by growth of seedlings on MS plates in air-tight containers in the dark supplied with either a flow of hydrocarbon-free air (Zero grade air, AirGas) or hydrocarbon-free air with 10 ppm (ppm) ethylene as previously described [28 (link)].
Zero grade air
Manufactured by Airgas
Zero grade air is a high-purity compressed gas used in laboratory applications. It is a mixture of nitrogen and oxygen, free of contaminants and impurities. Zero grade air meets stringent purity specifications to ensure consistent and reliable performance in sensitive analytical instruments and processes.
Automatically generated - may contain errors
4 protocols using zero grade air
1
Arabidopsis Seed Sterilization and Growth
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Arabidopsis seeds were surface-sterilized in 50% bleach with 0.01% Triton X-100 for 15 min and washed five times with sterile, doubly distilled H2O before plating on MS medium (4.3 g MS salt, 10 g sucrose, pH 5.7, 8 g phyto agar per liter). After 3-4 days of cold (4 °C) treatment, the plates were wrapped in foil and kept in at 24 °C in an incubator before the phenotypes of seedlings were analyzed. For propagation, seedlings were transferred from plates to soil (Pro-mix-HP) and grown to maturity at 22 °C under 16-h light/8-h dark cycles. Ethylene treatment of Arabidopsis seedlings was performed by growth of seedlings on MS plates in air-tight containers in the dark supplied with either a flow of hydrocarbon-free air (Zero grade air, AirGas) or hydrocarbon-free air with 10 ppm (ppm) ethylene as previously described [28 (link)].
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Arabidopsis seeds were surface-sterilized in 50% bleach with 0.01% Triton X-100 for 15 min and washed five times with sterile, doubly distilled H2O before plating on MS medium (4.3 g MS salt, 10 g sucrose, pH 5.7, 8 g phyto agar per liter). After 3-4 days of cold (4 °C) treatment, the plates were wrapped in foil and kept in at 24 °C in an incubator before the phenotypes of seedlings were analyzed. For propagation, seedlings were transferred from plates to soil (Pro-mix-HP) and grown to maturity at 22 °C under 16-h light/8-h dark cycles. Ethylene treatment of Arabidopsis seedlings was performed by growth of seedlings on MS plates in air-tight containers in the dark supplied with either a flow of hydrocarbon-free air (Zero grade air, AirGas) or hydrocarbon-free air with 10 ppm (ppm) ethylene as previously described [28 (link)].
2
Catalytic Hydrocarbon Reaction Study
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1-Butene (5.4 mol% in N2) and a GC calibration standard of 1-butene, cis-2-butene, trans-2-butene, 1,3-butadiene, and n-butane (1 mol% each in N2) were purchased from Matheson Trigas. A GC calibration standard for CH4, C2H4, C2H6, C3H6, C3H8, and 1-butene was purchased from Praxair. N2, CO2, and zero-grade air were purchased from Airgas. 1-Butanol (99%) and hydrated nitrate salts of Cr, K, Ca, Ni, Fe, Cu, and Ce were purchased from Sigma-Aldrich, as was the oxalate salt of Mo. The γ-Al2O3 catalyst support was from Sasol (Puralox Nwa-155). Inert quartz chips (nominally 30–50 mesh) were from Dupré Minerals.
3
Arabidopsis Seed Sterilization and Ethylene Treatment
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Arabidopsis seeds were surface-sterilized in 50% bleach with 0.01% Triton X-100 for 15 min and washed five times with sterile, doubly distilled H2O before plating on MS medium (4.3 g MS salt, 10 g sucrose, pH 5.7, 8 g phyto agar per liter). After 3–4 days of cold (4 °C) treatment, the plates were wrapped in foil and kept in at 24 °C in an incubator before the phenotypes of seedlings were analyzed. For propagation, seedlings were transferred from plates to soil (Pro-mix-HP) and grown to maturity at 22 °C under 16-h light/8-h dark cycles. Ethylene treatment of Arabidopsis seedlings was performed by growth of seedlings on MS plates in air-tight containers in the dark supplied with either a flow of hydrocarbon-free air (Zero grade air, AirGas) or hydrocarbon-free air with 10 ppm (ppm) ethylene as previously described [33 (link)].
4
Arabidopsis Ethylene Response Assay
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Arabidopsis seeds were surface-sterilized in 50% bleach with 0.01% Triton X-100 for 15 min and washed five times with sterile, doubly distilled H2O before plating on MS medium (4.3 g MS salt, 10 g sucrose, pH 5.7, 8 g phytoagar per liter) with or without addition of 10 μM 1-aminocyclopropane-1-carboxylic acid (ACC, Sigma), the biosynthetic precursor to ethylene. After 3–4 days of cold (4 °C) treatment, the plates were wrapped in foil and kept in at 24 °C in an incubator before the phenotypes of seedlings were analyzed. For propagation, seedlings were transferred from plates to soil (Pro-mix-HP) and grown to maturity at 22 °C under 16-h light/8-h dark cycles. Ethylene treatment of Arabidopsis seedlings was performed by growth of seedlings on MS plates in air-tight containers in the dark supplied with either a flow of hydrocarbon-free air (Zero grade air, AirGas) or 4 h hydrocarbon-free air with 10 p.p.m.) ethylene as previously described12 (link). For hypocotyl length measurements, 3-day-old seedlings were scanned using an Epson Perfection V700 Photo scanner, and hypocotyls were measured using NIH Image (http://rsb.info.nih.gov/nih-image/ ).
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