PreparationofYeastDNAEmbeddedinAgarosePlugs
Preparation of Yeast DNA Embedded in Agarose PlugsAnja van Brabant(adapted from Iadonato, S. P., and A. Gnirke. 1996. RARE-cleavage analysis of YACs. Methods Mol Biol 54: 75-85.)1. Inoculate a 5 ml culture with a single colony from a YAC-containing strain of yeast and grow until saturated. Determine the number of yeast cells per milliliter. The cell count should be roughly 1 X 108 cells/ml for a saturated c......阅读全文
QUALITATIVE－ANALYSIS－OF－DNA－FRAGMENTATION－BY－AGAROSE－GEL－ELECTROPHORESIS
1. IntroductionNuclear morphology changes characteristic of apoptosis appear within the cell together with a distinctive biochemical event: the endonu
DNA电泳（agarose胶）操作方法
一、 试剂与材料：1、 琼脂糖2、 电泳缓冲液（1×TAE）3、 10mg/ml溴化乙锭4、 上样缓冲液5、 电泳仪和水平电泳漕6、 透射紫外灯7、 胶带纸 二、 操作方法 按1～2%的琼脂糖浓度（据DNA样品分子量不同而定）配胶100ml ↓ 置于微波炉内加热搅拌，至琼脂糖完全溶解
Quick－and－Easy－Isolation－of－Genomic－DNA－from－Yeast
ProcedureTransfer 1.5 ml of liquid culture of yeast grown for 20 - 24 h at 30°C in YPD (1% yeast extract, 2% peptone, 2% dextrose) into a microcentrif
Restriction－Digests－of－High－Molecular－Weight－Yeast－DNA
Purpose:To perform restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion.Time req
Column－Method－for－Lambda－Phage－DNA－Preparation
Purpose:Mini-prep method for lambda phage DNA purification from lysates.Time required:4 hours once the lysate is in handSpecial supplies required:BioR
Construction－of－BAC－Libraries：Construction－of－a－BAC－library
Once high molecular weight (HMW) DNA has been prepared it must somehow be fragmented and DNA in the desired size range isolated. In general, as the de
组织学——显微解剖
Laser Capture Microdissection (LCM)Introduction to LCM (BJMU) Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections (NIH Laser Capture M
Standard－Operating－Procedures－for－T1Phage－Testing－Assay
I. Introduction:This assay uses a lawn of phage-susceptible E. coli (DH10B) embedded in a layer of agarose. This top agarose lays on a bed of standard
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Maxipreparation
实验概要Plasmid DNA is isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and SolutionsAlkaline lysis solu
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Minipreparation
实验概要Plasmid DNA is isolated from small-scale (1-2 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and Solutions: Alkaline lysis
Genomic－DNA－Extraction－－PureLink™
实验概要The PureLink™ Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently isolate g
包埋－Embedded－Samples
包埋 Embedded Samples可以将部分样品包埋在1-4%浓度的低熔点琼脂糖中，不同浓度的琼脂糖其稳定性不同，对活体样品的影响也不同。适合样品：斑马鱼、果蝇、线虫、拟南介等荧光标记的模式生物。包埋尺寸：平台配有不同规格的毛细管和1ml注射器，样品大小0.23mm-3.06mm，介于毛细管和注
DNA电泳
DNA电泳（主要内容如下） Preparation of Agarose Gel and Electrophoresis Extraction of DNA From Agarose Gel Extraction of DNA from Acrylamide Gels DNA Marker
QUALITATIVE－ANALYSIS－OF－DNA－FRAGMENTATION－BY－AGAROSE－GEL－ELECTROPHORESIS2
3. Commentary 3.1. Background informationApoptosis is an innate mechanism of eukariotic cell suicide which plays a major role in many physiological
酵母转化
· Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation· Yeast Transformation (Breeden Lab)LiAc method· Large-Scale Y
Cosmid－Cloning:－Cell－preparation,－DNA－packaging,－and－Cell－Transfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction man
A－practical－method－for－the－preparation－of－total－DNA－from－filamentous－fungi
Most methods of DNA preparation from fungi are time-consuming due to the need to first make protoplasts, expensive for chemicals such as cesium chlori
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
其它PCR方法
· Standard PCR Protocol (Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
Immunohistochemistry－on－paraffin－embedded－sections
主要试剂1. Neutral-buffered Formalin, 10% (NBF), 1 liter (Commercially available pre-prepared from many laboratory reagent suppliers).Double -distilled H
DNA克隆
DNA克隆（主要内容如下）· General Procedure· PCR Cloning· Subcloning· ET Cloning· Vector Preparation· Ligation Re
组织学——组织制备
· Histological techniques (William H. Heidcamp)Very detailed guide to histological techniques, like fixation, dehydration, embedment and subs
DNA微序列技术
· Protocols for Making Drosophila Arrays (Stanford U.)Detailed protocol for making arrays including PCR Amplification of cDNAs for Printing,
DNA琼脂糖凝胶电泳（agarose－gel－electrophoresis）分析
一、原理琼脂糖凝胶具有分子筛效应。在中性ppH值的电泳缓冲液体系中，DNA分子由于带负电荷，所以在电场作用下由负极向正极泳动。由于DNA分子的大小和构型不同，在相同的时间内迁移至不同的位置。凝胶经溴化乙锭染色后，紫外检测仪下观察，即可看见DNA片段按大小不同呈条带分布。由于在一定条件下，DNA的迁移
琼脂糖凝胶电泳（agarose－gel－electrophoresis）检测DNA
原理： 琼脂糖是从海藻中提取出来的一种线状高聚物，可作为电泳支持物，适用于分离大小范围在0.2-50kb的DNA片段。DNA分子的迁移率与分子量的对数值成反比关系。观察其迁移距离，与标准DNA片段进行对照，就可获知该样品分子量大小。在质粒抽提过程中，由于各种因素的影响，使质粒DNA呈现超螺旋的共
Yeast－Media
YEPD (non-selection)-1% yeast extract-2% peptone-1.5% agar (if needed for plates)After autoclaving add glucose to 2% by adding 100 ml of 20% solution
酵母遗传学技术
Genome-wide Gene Expression Analysis (Richard Young Research Group,Whitehead Institute for Biomedical Research)A genoe-wide gene expression analysis u
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA. EmbeddingB. CuttingC. StainingII. Pr