In vitro growth of seedlings

作者：Schneitz…    实验频道来源：本站原创    点击数：    更新时间：2003-11-2

sterilisation of seeds: rinse with 70% EtOH for 30 sec put in 1% bleach (sodium hypochlorite, supplemented by a few drops of Tween-20) for 5 min wash 2-4x with sterile H2O for 1 min plating and growth resuspend in sterile 0.1% agarose plate 4 ml in petridish (0.8% agarose supplemented with 1/2 MS10) in sterile hood let dry for 1-2 h in the hood wrap with 3M micropore tape keep at 4°C for 4 days place the dishes under light Solutions 1/2 MS10       MS Macro I 50.0 ml MS Macro II 50.0 ml Myo 10.0 ml FeNaEDTA 10.0 ml MS Vitamines 1.0 ml MS Micronutrients 0.5 ml Sucrose 10.0 g  H2O x.x ml Total 1000.0 ml adjust pH to 5.7 with 1 M KOH, alternatively MES (0.5g/l) can be used to ensure more stable pH during plant growth. Remarks It is essential that the medium is autoclaved in either a small autoclave or pressure cooker because prolonged exposure to heat will cause complex formation between sucrose and phytoagar which becomes visible as a yellow coloring of the medium. This medium will be softer after hardening than correctly autoclaved medium. The MS medium protocol is taken from Murashige and Skoog (1962) Physiologia Plantarum 15: 473-497. The recipe can be found here. Materials Reagent/Tool Supplier Cat.-#       Agarose Quantum-Appligene 130022 MS media Phytotechnology Laboratories   Petridish 145/20 mm Greiner Labortechnik 639 102 Sodium hypochlorite 13% Fluka 71696 Tween-20 Fluka 93773 3M Micropore tape Galenica 处理 SSI 文件时出错