DNA实验技术

DNA absorbs ultraviolet light due to its highly conjugated nature. DNA may thus be easily quantitated in a UV spectrometer.Typically 1 OD260 (i.e. a solution having an absorbance of one unit at 260 nm ...

Sample DNAs should not be highly degraded nor contain PCR inhibitors such as high concentrations of heme or chelating agents. For each individual assayed 250 ng of genomic DNA are digested separately ...

'Phenol' as used is Analar grade. Phenol should be melted at 65℃8-hydroxyquinoline added to a final concentration of 0.1% and equilibrated three times with an equal volume of 1M Tris.HCl pH 7.0. The f ...

Salmon Sperm DNA1.Dissolve 1 g salmon sperm DNA in 100 ml H2 O. 2.Autoclave (20 minutes) and aliquot in 1 ml/tube 3.Store at -20℃. (common freezer for stock solutions) ...

DNaseI FootprintintSolutions10X Binding Buffer200 mM Tris 8.0 200 m l 1M Tris pH 8.0500 mM NaCl 100 m l 5M NaCl10 mM EDTA 20 m l 0.5 M EDTA pH 8.0680 m l Qstore at room temperatureDNaseI Dilution Buff ...

Protocol For the original protocol describing CGH see the article by Kallioniemi et al . For an example of how this technique has been utilized by the NCI Prostate Cancer Working Group see the section ...

用于Genome Walking Kit的模板总DNA 一般要满足以下几点：1、DNA 的完整性，电泳为单一条带。2、避免杂质，OD260/280=1.8~2.0。3、模板DNA 不要被污染。4、准确定量，不要少于3μg。 ...

Protocols for HapMap assay-design Affymetrix platform (used by Broad)Defined protocols:LSID: urn:LSID:affymetrix.hapmap.org:Protocol:affy_assay_design_1:1Title: Genotyping using Affymetrix arraysDescr ...

An end labeled DNA probe is incubated with a purified DNA-binding factor or with a protein extract. The unprotected DNA is then digested with DNase I such that on average every DNA molecule is cut onc ...

FINGERPRINTING PROTOCOL (AGAROSE GEL) * Restriction digests consist of:15.75 ml ddH2 O 1 µl 10 X buffer B (Boehringer Mannheim) 0.25 µl HindIII (40 U/µl) 3 µl DNA Set up digestions in 96 well plates. ...

Whole-Genome and Custom Fine-Tiling Array CGHComparative Genomic Hybridization (CGH) measures DNA copy number differences between a reference genome and your sample genome. NimbleGen offers two high-d ...

点击浏览该文件Footprinting Analysis ...

1. For double-stranded DNA templates:a. Denature template: 10ml DNA (5-10g or alkaline lysis mini prepDNA)8ml ddH2O　　2ml 2N NaOH　　-incubate 30' at 37℃.b. Dry-ice precipitate: 10ml 4M NH4OAc100ml EtOH　 ...

Buffers and gel solutions Long Ranger: we started using this in early 1995. Great stuff; the best thing is that the gels are not sticky after drying even without removal of the urea. Long Ranger Gel S ...

Miniprep of BAC DNA1.Inoculate 4 - 8 tubes of 3ml/tube LB with 15μg/ml chloramphenicol with BAC clone and grow oveernight. Miniprep with AutoGen 740 according to the protocol described in our Web p ...

单拷贝克隆产量低，高拷贝克隆稳定性差，一直让研究者在克隆 表达时难以选择。蛋白的表达就有诱导表达系统，可以实现人为地控制蛋白的表达时间和表达量——现在连质粒的拷贝数可以借助诱导的方法来人为控制了！Epicentre的专利技术——CopyControl克隆 系统能够让您共享单拷贝和高拷贝的优点。CopyControl克隆，首先以单拷贝形式生长——单拷贝可以确保插入稳定及成功克隆，编码毒性基因序列—— ...

一、 目的及要求。1、 了解质粒作为载体在基因工程中的作用2、 熟记提取质粒的基本原理，学习提取过程和方法3、 为下一步实验提供高纯度的DNA 样品二、原理：1、 质粒（Plasmid）：独立于染色体以外的双链、闭合、环状DNA，可自我复制。为基因工程中的常用载体（Vector）2、 作为载体的质粒需具备以下特点：1） 能自主复制。2） 具有多个限制性内切酶的单一酶切位点，又称为多克隆位点（mul ...

【实验目的】掌握植物总DNA的抽提方法和基本原理。学习根据不同的植物和实验要求设计和改良植物总DNA抽提方法。【实验原理】通常采用机械研磨的方法破碎植物的组织和细胞，由于植物细胞匀浆含有多种酶类(尤其是氧化酶类)对DNA的抽提产生不利的影响，在抽提缓冲液中需加入抗氧化剂或强还原剂(如巯基乙醇)以降低这些酶类的活性。在液氮中研磨，材料易于破碎，并减少研磨过程中各种酶类的作用。十二烷基肌酸钠(sark ...

Q:my group is involved in a national project to develop a genomic platform for cancer research. while Nimblegen has developed target enrichment chip for the Genome Sequencer 20 nothing similar has bee ...

【实验目的】（1）学习并掌握动物组织总DNA的提取方法及其原理。（2）从肝脏组织中提取到一定量的纯净的DNA样品。【实验原理】DNA是一切生物细胞的重要组成成分，主要存在于细胞核中。通过研磨和SDS作用破碎细胞；苯酚和氯仿可使蛋白质变性，用其混合液（酚：氯仿：异戊醇）重复抽提，使蛋白质变性，然后离心除去变性蛋白质；RNase降解RNA，从而得到纯净的DNA分子。【仪器、材料、试剂】（一）仪器1．高 ...