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Dynabeads® mRNA DIRECT Kit

实验原理 The isolation protocol relies on base pairing between the polyA residues at the 3’ end of most mRNA and the oligo (dT)25 residues covalently coupled to the surface of the Dynabeads® . Other RNA ...

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逆转录-聚合酶链式反应(RT-PCR)

实验试剂 1.RNA提取试剂2.第一链cDNA合成试剂盒3.dNTPmix:含dATP、dCTP、dGTP、dTTP各2mM4.Taq DNA聚合酶 实验步骤 1. 总RNA的提取:见相关内容。2. cDNA第一链的合成:目前试剂公司有多种cDNA第一链试剂盒出售,其原理基本相同,但操作步骤不一。现以GIBICOL公司提供的SuperScriptTM Preamplificat ...

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Growing Cells in Geltrex Reduced Growth Factor Basement Membrane Matrix

实验步骤 There are many applications for Basement Membrane Matrix which require different thicknesses and concentrations. In general a protein concentration 9 mg/ml is used for differentiation studies o ...

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DNA Probe Purification Protocol

实验试剂 Materials Supplied By User1. Microcentrifuge capable of at least 10000 x g.2. Sterile 1.5 mL centrifuge tubes.3. Sterile deionized water (or TE buffer)4. Absolute (or 95%) ethanol 5. Protective ...

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Organotypic Collagen I Assay: A Malleable Platform to Assess Cell Behaviour in a 3-Dimensional Context

实验材料 NameCompanyCatalog NumberComments10x MEMGibco21430-NaOHSigma367176-500GPrepare 0.22 M stock in waterFCSPAA LaboratoriesA15-101-35 mm dishesFalcon353001For step 3.460 mm dishesFalcon353004For st ...

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Isolation of CD34+ Cells from Mononuclear Cells (MNCs) from Human Bone Marrow, Peripheral Blood or Cord Blood

实验试剂 MagnetMixer allowing both tilting and rotationBuffer: PBS (without Ca2 and Mg2 ) w/0.1% BSA and anticoagulant pH 7.4DNase I (when required)Culture Medium: e.g. RPMI 1640 with 2% Fetal Calf Seru ...

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黑豆蚜胞内共生菌对日本柄瘤蚜茧蜂发育和繁殖的影响

实验材料 黑豆蚜用蚕豆苗饲养,日本柄瘤蚜茧蜂的日常饲养是在蚜虫2-3龄时接蜂,接蜂时间24小时。温度23-25℃,光周期16:8。 实验步骤 1. 蚜虫共生菌的脱除日常饲养的黑豆蚜是有共生细菌的蚜虫,称之为“共生蚜虫”,其胞内共生细菌的脱除采用饲喂抗生素利福平的方法。经利福平处理后,脱去共生细菌的蚜虫称为“脱共生蚜虫”。2. 蚜茧蜂在共生和脱共生蚜虫体内生长发育和繁殖的观察共生和脱共生 ...

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Fungal Midi DNA Kit Protocol for Fresh/Frozen Specimens

实验试剂 1. Equilibrate sterile dH2O water or DNA Elution Buffer at 65°C.2. Isopropyl alcohol (isopropanol)3. Absolute (96%-100%) ethanol 4. RNase A stock solution at 20 mg/mL 实验设备 1. Centrifuge cap ...

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Dynabeads® CD3CD28 CTS

实验原理 Dynabeads® CD3⁄CD28 CTS™ offer a simple method for isolation activation and expansion of human T cells. Firstly CD3 T cells are separated and concentrated from the apheresis product by ma ...

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植物基因组DNA的RFLP

实验原理 DNA多态性是指DNA碱基顺序的差异性。这种差异性不仅存在于不同的植物之间,也存在于同一种植物的不同个体之间。RFLP多态性是指DNA限制性内切酶酶切片段长度的多态性(restniction fragment length olymophorphism)。由于碱基顺序的差异,在不同的种或同一个种的不同个体之间,它们的基因组DNA限制性内切酶位点的种类和数目不同,经特定的限制性内切酶切割 ...

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Sandwich ELISA Protocol

实验试剂 Carbonate Coating Buffer8.4 g NaHCO33.56 g Na2CO3Add ddH2O up to 1.0L pH to 9.5Phosphate Buffered Saline (PBS):80.0 g NaCl14.4 g Na2HPO42.4 g KH2PO42.0 g KClAdd ddH2O up to 10 L pH to 7.2 with ...

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E.Z.N.A.® Viral RNA Spin Protocol

实验试剂 1. Absolute ethanol ( 96-100%) 实验设备 1. Sterile RNase-free pipette tips and microcentrifuge tubes2. Table top microcentrifuge at room temperature.3. 2-mercaptoethanol4. Disposable latex glov ...

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Silver Enhancement Protocol

实验试剂 Prepare the following reagents fresh daily except for the citrate buffer. 1. Citrate buffer: Dissolve 23.5 g trisodium citrate dihydrate and 25.5 g citric acid monohydrate in 850 ml distilled wa ...

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RNA isolation protocol: cells in culture

实验步骤 Procedure1.Using at least 106 cells aspirate off the media and wash X1 with ice cold PBS (1-2 ml)2.Aspirate off the PBS (Remove as much as possible) and add 1 ml of trizol3.Scrape the plate b ...

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RNA Purification Protocol for 100-200 mg of Animal Tissue

实验原理 E.Z.N.A.® SQ Tissue RNA uses a highly efficient solution based system to provide a convenient fast reliable and non-toxic method to isolate high quality RNA from various samples. Samples are fir ...

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测定植物体内可溶性蛋白质含量(LoWry法与劳里法)

实验原理 LoWry法是双缩脲法(BIureT)和福林酚法(FolIn-酚)的结合与发展。其原理是蛋白质溶液用碱性铜溶液处理后,碱性铜试剂与蛋白质中的肽键作用产生双缩脲反应,形成铜—蛋白质的络合盐。再加入酚试剂后,在碱性条件下,这种被作用的蛋白质上的酚类基团极不稳定,很容易还原酚试剂中的磷钨酸和磷钼酸(PHosPHoMolyBdATe & PHosPHoTungsTATe),使之生成磷钨蓝和磷钼 ...

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DD-PCR

实验试剂 T12G、T12C、T12A、10-mer的随机引物、RT-PCR试剂盒、测序胶、银染系统 实验设备 PCR仪、电泳仪、测序槽、银染设备 实验步骤 1. 总RNA提取(略)2. 残留DNA的去除反应体系 1-5ugRNA 10×DNA酶缓冲液 2ul RNA酶抑制剂 1ul ...

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Preparation of Powdered Tissue Culture Media

实验步骤 1.Remove Penicillin-Streptomycin solution and histidine solution (1 tube/ 500 ml) from -20 freezer to thaw.2.To a flask as close to the final volume of the media you wish to prepare add approx ...

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基因枪轰击洋葱表皮观察GFP瞬时表达

实验试剂 高渗培养液(MS无机盐、40g/L甘露醇),高渗固体培养基(MS无机盐、40g/L甘露醇、0.7% agar),无水乙醇,无菌去离子水,12.5 M CaCl2,0.1 M亚精胺,等渗培养基(MS培养基) 实验设备 高速离心机,1.5 ml Ep管,涡旋器,超净台,基因枪,气瓶,激光共聚焦显微镜 实验材料 洋葱内表皮 实验步骤 1. OsAGAP瞬时表达载 ...

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免疫共沉淀试剂配制及技术路线

实验试剂 1. RIPA Buffer配制基础成分: Tris-HCl(缓冲液成分,防止蛋白变性) NaCl(盐份,防止非特异蛋白聚集) NP-40(非离子去污剂,提取蛋白;用H2O配制成10%储存液) 去氧胆酸钠(离子去污剂,提取蛋白;用H2O配制成10%储存液;避光保存) 注意:准备激酶(致活酶)实验时,不要加去氧胆酸钠,因为离子型去污剂能够使酶变性,导致活性丧失。 RIPA蛋白酶抑制剂 苯 ...

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