DNA提取与纯化

Sample Preparation Cheek cells are obtained by rinsing the mouth with 25mls of any commercial mouth wash solution available for about 30sec the first thing in the morning. It is important not to brus ...

1.Grind in mortar and pestle or Waring blender with 5-7 volumes buffer A per g tissue. Use MCE at 350 l/L and if necessary with 5 ml 1 M DIECA/L. 2.Squeeze through cheesecloth two layers of Miracloth ...

Purpose: Purification of oligonucleotides (which have already been purified by reverse phase) can increase the efficiency of site directed mutagenesis from around 30% to ~75% or greater in some cases ...

细胞内的核酸多以核蛋白的形式存在，其中脱氧核糖核蛋白主要存在于细胞核中，核糖核蛋白主要存在于细胞质中。这两类核蛋白在 0.14mol /L 氯化钠溶液中的溶解度相差很大，核糖核蛋白在此溶液中具有很高的溶解度，脱氧核糖核蛋白的溶解度却相当低。制成肝匀浆后，用 0.14mol /L 氯化钠溶液抽提，可将两种核蛋白分离。分离过程中加入少量柠檬酸钠，可抑制脱氧核糖核酸酶对DNA 的水解作用。 SDS( ...

This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure with a variety of Populus species as well as tobacco and ...

概 述 基因组DNA的提取通常用于构建基因组文库、Southern杂交(包括RFLP)及PCR分离基因等。利用基因组DNA较长的特性，可以将其与细胞器或质粒等小分子DNA分离。加入一定量的异丙醇或乙醇，基因组的大分子DNA即沉淀形成纤维状絮团飘浮其中，可用玻棒将其取出，而小分子DNA则只形成颗粒状沉淀附于壁上及底部，从而达到提取的目的。在提取过程中，染色体会发生机械断裂，产生大小不同的片段，因此 ...

质粒是存在于细菌染色体外的一个或多个能独立复制并稳定遗传的小型环状双链DNA分子，其分子量一般在0. 2～10kD 范围内。由于质粒分子小，便于分离和提取，可以携带目的基因进入细菌、动物细胞或植物体内进行扩增与表达。基因克隆实验中常将经改造后的质粒作为运送基因的载体，质粒D2NA提取效率及质量对后续实验步骤(如酶切、连接、转化大肠杆菌与PCR扩增等)的成功与否有着直接的关系，因而高效快速地从细菌细 ...

Materials: Silica Suspension: add 2 g of silica to 15 ml of H2O wash 3x by centifugation at 2000 x g for 2 min estimate vol of silica and resuspend in 2 vol H2O Silica Wash Solution: 50 mM NaCl 10 mM ...

Tissue collection storage microdissection sectionin g: See separate protocol. Tissue handling : Note that all fresh tissue shoμld be handled as BioSafety Level 2 materials (wear gloves lab coat etc ...

摘　要　对植物中DNA抽提是在分子水平上对植物进行系统分析与研究的基础性工作.经多次实验，使用改良的CTAB法抽提山茶属植物中总DNA，即在抽提前加入抗氧化剂β-巯基乙醇，并使用冷冻的异丙醇来沉淀DNA， 获得了较好的效果，其纯度和得率完全能满足常规分子生物学操作的要求. 关键词　山茶属植物；DNA抽提；叶片；改良CTAB法 中图法分类号　Q523；Q949.758.4 DNA Ex ...

(adapted from Bruce A. Roe Department of Chemistry and Biochemistry The University of Oklahoma Norman Oklahoma 73019 broe@ou.edu) Typically 2.5 - 3 volumes of an ethanol/acetate solution is added to t ...

1. Transfer plugs of mycelial margin to 0.1 P plates. Allow to grow to fill plate (about one week at 21°). 2. Macerate two plates of mycelia in a blender with 50 ml of .002 P liquid medium. Inocul ...

- make a 5µm section to do an evaluation of the % tumour cells - make 50X50µm sections for the DNA isolation - put sections in a falcon tube containing 10 ml of 1XSE - add 100µg/ml p ...

Electroelution of agarose fragments Electroelution buffer 1 M Tris pH 7.5 12.0 mls 0.5 M EDTA 0.24 mls 1 M NaCl 3.0 mls qs to 600 mls dH2O Acetate cushion 3 M NaAcetate pH 4.8 480 μl 0.1 % Bromphe ...

Mitochondrial DNA Isolation from Somatic Embryogenic Cell Cultures of Larix (Excerpt from Thesis). Mitochondrial DNA is isolated by a modification of the methods described by Wilson and Chourey (1984) ...

A practical method for the preparation of total DNA from filamentous fungi M.I. Borges M.O. Azevedo R. Bonatelli Jr. M.S.S. Felipe and S. Astolfi-Filho Departamento de Biologia Celular Universidade de ...

SDS 高盐法(方案1) 　 具体步骤: 称取1g 土壤放入研钵中倒入适量的液氮立即研磨;再倒入适量的液氮研磨重复3 次使土壤颗粒研成粉末; 将13.5 ml 提取缓冲液(0.1 mol/L磷酸盐 0.1 mol/L EDTA 0.1 mol/L Tris-HCl 1.5 mol/L NaCl 1.0 % CTAB) 和50μl蛋白酶K(10 g/L ) 与5 g 土壤置于50 m ...

Abstract: Single strand conformational polymorphism (SSCP)is the most widely used PCR-based methods for point mutation detection. The abnormal band found by SSCP analysis is normally verified ...

原理 DNA 以核蛋白形式存在于细胞核中，制备 DNA 的原则是既要将 DNA 与蛋白质、脂类和糖类等分离，又要保持 DNA 分子的完整。蛋白酶K 及链霉蛋白酶 E 的应用使这两个原则得到了保证。 蛋白酶K 或链霉蛋白酶 E 均为广谱蛋白酶，其重要特性是能在 SDS 和 EDTA 存在下保持很高的活性。在匀浆后提取 DNA 的反应体系中， SDS 可破坏细咆膜、核膜，并使组织蛋白与 DNA 分子分 ...

Preparation of Sonicated Salmon Sperm DNA Procedure: 1) Use Pharmacia #27-4564-01. With clean flamed scissors and forceps weigh 0.25 g/50 ml conical and add 50 ml/conical of 0.02 M Tris pH 7.6. Allo ...