Acidobacteria strains were tested for their ability to solubilize a mineral form of phosphate. P. putida IAC-RBal4 was used as a positive control. Tests were performed on the National Botanical Research Institute’s phosphate growth medium (NBRIPM) containing per liter 15 g agar–agar ultrapure (Merck KGaA), 10 g glucose, 5 g Ca3(PO4)2, 5 g MgCl2·6H2O, 0.25 g MgSO4·7H2O, 0.2 g KCl and 0.1 g (NH4)2SO4 (Nautiyal 1999 (link)). All strains were inoculated by transfer from the 0.1 × TBA pH 5.0 media using inoculation loop and incubated for 6 weeks at 20 °C. The clearing zones around the colonies indicated phosphate solubilization by the isolates. The experiment was carried out in triplicate on separate plates.
Agar agar ultrapure
Manufactured by Merck Group
Agar-agar ultrapure is a highly purified form of agar, a natural gelling agent derived from red seaweed. It is a neutral, odorless, and colorless polysaccharide that forms a firm, transparent gel when dissolved in water and cooled. The core function of agar-agar ultrapure is to serve as a solidifying and gelling agent in various laboratory applications.
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3 protocols using agar agar ultrapure
1
Phosphate Solubilization by Acidobacteria
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Acidobacteria strains were tested for their ability to solubilize a mineral form of phosphate. P. putida IAC-RBal4 was used as a positive control. Tests were performed on the National Botanical Research Institute’s phosphate growth medium (NBRIPM) containing per liter 15 g agar–agar ultrapure (Merck KGaA), 10 g glucose, 5 g Ca3(PO4)2, 5 g MgCl2·6H2O, 0.25 g MgSO4·7H2O, 0.2 g KCl and 0.1 g (NH4)2SO4 (Nautiyal 1999 (link)). All strains were inoculated by transfer from the 0.1 × TBA pH 5.0 media using inoculation loop and incubated for 6 weeks at 20 °C. The clearing zones around the colonies indicated phosphate solubilization by the isolates. The experiment was carried out in triplicate on separate plates.
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Acidobacteria strains were tested for their ability to solubilize a mineral form of phosphate. P. putida IAC-RBal4 was used as a positive control. Tests were performed on the National Botanical Research Institute’s phosphate growth medium (NBRIPM) containing per liter 15 g agar–agar ultrapure (Merck KGaA), 10 g glucose, 5 g Ca3(PO4)2, 5 g MgCl2·6H2O, 0.25 g MgSO4·7H2O, 0.2 g KCl and 0.1 g (NH4)2SO4 (Nautiyal 1999 (link)). All strains were inoculated by transfer from the 0.1 × TBA pH 5.0 media using inoculation loop and incubated for 6 weeks at 20 °C. The clearing zones around the colonies indicated phosphate solubilization by the isolates. The experiment was carried out in triplicate on separate plates.
2
Arabidopsis Growth Conditions for Epigenetic Studies
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For the training set, wild-type (WT) Col-0 and ddm1-2 [43 (link)] plants were grown in vitro on solid GM medium (MS salts (Duchefa), 1% sucrose, 0.8% Agar-agar ultrapure (Merck), pH 5.8) in a culture chamber under a 16 h light (light intensity ∼150 μmol·m−2·s−1; 21 °C) and 8 h dark (19 °C) photoperiod.
For the Dark/Light set, seeds from wild-type (WT) Col-0 arabidopsis plants were surface-sterilized, plated on filter papers lying on MS medium supplemented with 0.9% agar and exposed to either a 16-/8-h (23/19 °C) white light/dark photoperiod or constant dark conditions (wrapped in 3 layers of aluminum foil). White light is generated by fluorescent bulbs (100 μmol·m−2·s−1). Seedlings are harvested under light condition or under safe green light for the dark condition [35 (link)].
For the Dark/Light set, seeds from wild-type (WT) Col-0 arabidopsis plants were surface-sterilized, plated on filter papers lying on MS medium supplemented with 0.9% agar and exposed to either a 16-/8-h (23/19 °C) white light/dark photoperiod or constant dark conditions (wrapped in 3 layers of aluminum foil). White light is generated by fluorescent bulbs (100 μmol·m−2·s−1). Seedlings are harvested under light condition or under safe green light for the dark condition [35 (link)].
3
Arabidopsis Mutant Cultivation Protocol
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The Arabidopsis thaliana wild-type (WT); ddb2-3 [23 ], uvr3 phrI (WiscDsLox334H05 and WiscDsLox466C12, [39 (link)]), nrpd1 (Salk_583051), nrpe1 (Salk_029919), dcl2-1 (Salk_064627), dcl3-1 (Salk_005512), dcl4-2 (GABI_160G05), ago1-27 [42 ] plants used in this study are in the Columbia ecotype (Col0). Plants were grown in vitro on solid GM medium [MS salts (Duchefa), 1% sucrose, 0.8% Agar-agar ultrapure (Merck), pH 5.8] in a culture chamber under a 16 h light (light intensity ∼150 μmol m−2 s−1; 21°C) and 8 h dark (19°C) photoperiod.
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