============================================================================ medium no. 53: modified Gamborgʼs B5 medium, 0.25% (w/v) gellan gum, pH 5.7 ============================================================================ Plant cell lines ================= * :doc:`/cell_lines/rpc00067` Chemicals and stock solutions ============================== (All stock solutions are stored at 4°C) * B5 salt mix Gamborgʼs B5 Medium Salt Mixture, FUJIFILM Wako Pure Chemical Corporation (#399-00621) * Sucrose * MS_VT .. csv-table:: :header: "Chemical", "Concentration (mg/mL)" :widths: auto "Nicotinic acid", "0.5" "Pyridoxine·HCl", "0.5" "Thiamine·HCl", "0.1" "Glycine", "2" * MS_inositol .. csv-table:: :header: "Chemical", "Concentration (mg/mL)" :widths: auto "*myo*-Inositol", "40" * Gellan gum * KOH (1 N) Medium preparation =================== #. Dissolve the following chemicals in approximately 800 mL of distilled water. .. csv-table:: :header: "Chemical", "Amount" :widths: auto "B5 salt mix", "1 bag (1 L)" "Sucrose", "20 g" #. Add following stock solutions, and fill up to approximately 950 mL with distilled water. .. csv-table:: :header: "Stock solution", "Volume (mL)" :widths: auto "MS_VT", "1" "MS_inositol", "2.5" #. Adjust the pH of the solution to 5.7 with KOH (1 N), and fill up to 1 L with distilled water. #. Add 2.5 g of gellan gum to the medium. #. Autoclave the medium at 121°C for 20 min. Formulation ============ .. csv-table:: :header: "Chemical", "Concentration (mg/L)" :widths: auto "KNO\ :sub:`3`", "2500" "(NH\ :sub:`4`\ )2SO\ :sub:`4`", "134" "CaCl\ :sub:`2`\ ·2H\ :sub:`2`\ O", "150" "MgSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "250" "NaH\ :sub:`2`\ PO\ :sub:`4`\ ·H\ :sub:`2`\ O", "150" "H\ :sub:`3`\ BO\ :sub:`3`", "3" "MnSO\ :sub:`4`\ ·H\ :sub:`2`\ O", "10" "ZnSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "2" "KI", "0.75" "Na\ :sub:`2`\ MoO\ :sub:`4`\ ·2H\ :sub:`2`\ O", "0.25" "CuSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "0.025" "CoCl\ :sub:`2`\ ·6H\ :sub:`2`\ O", "0.025" "FeSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "27.8" "Na\ :sub:`2`\ -EDTA", "37.3" "Nicotinic acid", "0.5" "Pyridoxine·HCl", "0.5" "Thiamine·HCl", "0.1" "Glycine", "2" "*myo*-Inositol", "100" "Sucrose", "20000" "Gellan gum", "2500"