水稻转化培养基组份

水稻转化培养基组份N6D2:1)N6NutrientStockN6大量(20X)N6微量(20X)2)IronStock(100X)3)VitamineStock烟酸1mg/mlVB11mg/mlVB61mg/mlGlycine2mg/ml肌醇50mg/ml4)Sucrose5)2,4-D2mg/ml6)CaseinEnzymaticHydrolysat7)Phytagel1LpH5.850ml50ml10ml0.5ml0.1ml0.5ml1ml2ml30g1ml1g/l2.0—2.5g/lN6D2C(forco-culturing)N6D2mediumGlucoseAS(20mg/ml)10g/lto20mg/lfinalconc.N6D2S1(forselecting)NDmedium62HygromycinCefotaximineto25mg/lfinalconc.to300—400mg/lfinalconc.(现用300mg/l)N6D2S2(forselecting)NDmedium62HygromycinCefotaximineto50mg/lfinalconc.to200mg/lfinalconc.预分化200mlddH2O中加入0.5gphytagel,灭菌再依次加入HigrowRiceMedium50XSaltsI,II,III各4ml,微波炉加热消毒灭菌,加入hygromycin50mg/l,Cefotaximine200mg/l.分化培养基MSRCHpH5.81)MS0.25g/l水解酪蛋白CH(可不加)Sucrose30g/lphytagel2.5g/l激素终浓度6-BA2mg/lNAA0.2mg/lZeatin0.2mg/lKT0.5mg/lHygromycin50mg/lCefotaxime200mg/lTOC\o"1-5"\h\z*MS培养基 1LMS大量(20X) 50mlMS微量(100X) 10mlFe-EDTA(100X) 10mlVitamine烟酸0.5mg/lVB60.5mg/lVB10.1mg/lGlycine2mg/l肌醇100mg/lpH5.81/2MS培养基(生根培养基) 1LpH5.81.1/2MS盐分25ml5ml5ml1mg/l25ml5ml5ml1mg/l0.5mg/l30g/l2.g50mg/lMS微量(100X)Fe-EDTA(100X)2.MS维生素3.多效唑NAASucrosePhytagelHygromycineAAM培养基1LpH5.28磅灭菌1.AA大量(20X)50ml2.AA微量(100X)10ml3.Fe-EDTA(100X)10ml4.Vitamine烟酸0.5mg/lVB60.5mg/lVB11mg/lGlycine2mg/l肌醇100mg/l5.AAL-Gln880mg/lL-Asp270mg/lL-Arg230mg/lL-Gly80mg/l6.CH（水解酪蛋白）500mg/l可不加7.Sucrose68.5g/l8.Glucose36g/lAB1LpH7.28磅灭菌AB大量(20X)50mlAB微量(100X)10mlFe-EDTA(100X)1mlGlucose(无水)5g微量:MnSO4.H2O微量:MnSO4.H2OZnSO4.7H2OH3BO3KINutrientStock:(20X)Stockconc.Finalconc.大量:KNO356.6g/l2.83g/l(NH4)2SO49.26g/l463mg/lKH2PO48.0g/l400mg/lMgSO4.7H2)3.7g/l185mg/lCaCl2.2H2O3.3g/l 166mg/l66mg/l 3.3mg/l30mg/l 1.5mg/l32mg/l 1.6mg/l16mg/l 0.8mg/l*IronStock(100X)FeSO4.7H2ONa2EDTA2.8g/l 28mg/l3.7g/l 37mg/lMacronutrients(20x):g/L AA MSABKH2PO43.43.48KNO3--3856.6(NH4)2SO4----9.26nh4no3--33--MgSO47H2O7.47.43.7*CaCl.2HO228.88.83.32KC158.8----KHPO,60NaH2PO420NHCl420*独立溶解后加入。Micronutrients(100x):mg/L AA MSABMnSO.HO4 216901690250ZnSO.7HO4 2860860180H3BO3620620160CuSO.5HO4 22.52.5--NaMoO.2HO2 4 22525--CoCi.6HO222.52.5--KI838383MgSO4.7H2O30gKCl15gCaCl2lg

实验内容培养基及试剂实验器材诱导愈伤组织ND，无菌水，NaClO,吐62温-20,75%乙醇无菌大滤纸，镊子，解剖刀，药匙，无菌二角瓶转化及共培养AB，AAM（悬菌时加乙酰丁香酮至20mg/l）,N6D2C（滤纸用含20