CosmidCloning:Cellpreparation,DNApackaging,andCellTransfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction manualHost bacteria (E. coli LE392)preparation:Pick one colony from fresh overnight culture on NZY agar plate and inoculate 50 ml NZY broth supplemented with 0.2% maltose and 10 mM MgSO4.Grow at 37C, shaking, 4-6 h (do not grow past OD600 1.0/ml).Pellet bacteria 2000 rpm f......阅读全文
Cell子刊：“垃圾”DNA的阴暗面
在基因之间的DNA片段，散布着重复序列，曾经被认为是“基因组垃圾”，但现在科学家们了解到，一些这样的垃圾DNA并不是无害的。在《Cell Reports》发表的一项研究中，来自北卡罗来那大学（UNC）Lineberger综合癌症中心的研究人员报道称，某些短的DNA重复序列，或“垃圾DNA”，在尤文肉
Cell：新方法加速揭示致病“垃圾”DNA
由来自德国、美国和挪威等国家的科学家们组成的一个国际研究小组，在1月16日的《细胞》(Cell)杂志上发表论文描述了他们开展的一项新研究，该研究的目的在于更好地了解遗传变异使得某些个体易患诸如2型糖尿病等疾病的机制。结合他们的计算方法和实验，新研究解析并验证了2型糖尿病的潜在遗传病因。大体上，这
Cell子刊：癌症患者独有的古老DNA
一种改变了尼安德特人(Neanderthal)和丹尼索瓦人(Denisovan)的古老逆转录病毒，现在被发现同样也在现代人类——某些癌症患者的DNA中留下了一些改变。来自英国的一个研究人员小组在《当代生物学》(Current Biology)杂志的一篇论文中，著述了这一惊人的研究发现。
Cell惊人发现：DNA复制方案因人而异
人类细胞每次分裂的时候，都得复制六十亿DNA碱基，一个一个来显然是不现实的。事实上，DNA复制机器会同时介入多个起始点，进行分工合作。 哈佛大学医学院、Broad研究所和MIT的科学家们发现，人与人之间的DNA复制方案并不相同。他们鉴定了首个调控DNA复制时序（Replication timi
Mitochondrial－DNA－Isolation－from－Somatic－Embryogenic－Cell－Cultures－of－Larix
Mitochondrial DNA is isolated by a modification of the methods described by Wilson and Chourey (1984) and Radetzky (1990). Cell cultures at four days
定量PCR实验技术－QPCR
Quantitative PCRJoseph SambrookPeter Maccallum Cancer Institute and The University of Melbourne, AustraliaDavid W. RussellUniversity of Texas Southwes
siRNA－转染程序
A．siRNA转染的方法 哺乳动物转染的常见方法有：磷酸钙共沉淀、电穿孔法、DEAE-葡聚糖和polybrene、机械法（例如,显微注射和基因枪）、阳离子脂质体试剂，其中阳离子脂质体试剂转染法是目前最常用的转染方法。应用脂质体型转染试剂进行转染需要注重的几个方面：1. 转染试剂的用
超级感受态细胞的制备
The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" CellsJoseph SambrookPeter Maccallum Cancer Institute and T
Combined－Flow－Cytometric－Measurement－of－Two－CellSurface－Antigens
INTRODUCTIONFlow cytometry is frequently used to assess nucleic acid content in individual cells. Based on DNA content alone, however, cells in the qu
酵母转化
· Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation· Yeast Transformation (Breeden Lab)LiAc method· Large-Scale Y
PCR的下游应用
・ Agarose Gel Electrophoresis of PCR Products(Robert H. Cruickshank)・ Agarose Gel Electrophoresis of PCR Products(Immunology Resource)
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
Preparation of Luciferin forIn Vitro Bioluminescent AssaysMaterials• D-Luciferin Firefly, potassium salt, 1.0 g /vial(Caliper Life Sciences Part Numbe
Preparation－of－phage－particles－from－phage－vectors
Pick up one phage vectors-containing colony with a sterile loop and put into 10 ml 2xTY + 10 µg/l tetracycline.Shake at 200 rpm and 37 °C untill the
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
实验概要Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological
Preparation－Of－Ciliated－Protozoa－For－Scanning－Electron－Microscopy
Preparation Of Ciliated Protozoa For Scanning Electron MicroscopyGeneral notes: The same procedures are used to fix and stain cells for SEM and for TE
Preparation－of－bOGDOPG－mixed－micelles
Materials:All glassware must be acid washed, rinsed thoroughly with water then rinsed with acetone and dried.Redistill acetone.Recrystallizing BOG:1)
Preparation－Of－Peripheral－Blood－Cells－For－Chromosome－Analysis
实验概要Lymphocytes are differentiated cells which normally do not undergo subsequent cell divisions. By culturing lymphocytes in the presence of a mito
Adrenal－chromaffin－granule－(chromaffin－vesicle)－preparation
Adrenal chromaffin granule (chromaffin vesicle) preparationIntroduction. This prep is adapted from the classic paper of Smith and Winkler (Smith AD; W
TEM－Specimen－Preparation：Preparative－Techniques－for－the－TEM
For routine transmission electron microscopy (TEM), it is generally accepted that specimens should be thin, dry and contain molecules which diffract e
Preparation－of－Conventional－Actin－from－Skeletal－Muscle
Modified from Spudich & Watt, 1971, JBC 246:4866.1. Mix 20 ml buffer G with each gm of muscle acetone powder. Extract with stirring on ice for 30 min.
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
实验概要Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological
Preparation－of－fixed－embryos－for－immunocytochemistry－and－AP－staining
1. Transfer 50 ml of embryo cultures to centrifuge tubes. Spin at 1500 rpm for 5 minutes. Check that you can see a pellet of embryos at the bottom.Qui
In－Vivo－Imaging－of－Far1
In Vivo Imaging of Far-red Fluorescent Proteins after DNA Electrotransfer to Muscle TissueDNA electrotransfer to muscle tissue yields long-term, high
Heterogeneity－of－SingleCell－Gene－Expression－Across－Phenotypically（一）
Introduction Multi-cellular populations are fundamentally driven by the collective properties of individual cells. However, our understanding of ge
DNA纯化手册1
Purification of plasmid DNA (miniprep) with high yields using diatomaceous earthKyung-Soo Kim and Charles K. PallaghySchool of Botany, La Trobe Univer
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends01
IntroductionThe following protocol describes a procedure for the purification and cloning of miRNAs and other small RNAs in the 20-30 nucleotide size
如何有效地降低PepMuteTM，GenMuteTM－and－PepMuteTM－Plus－siRNA转...
如何有效地降低PepMuteTM，GenMuteTM and PepMuteTM Plus siRNA转染试剂的毒性PepMuteTM，GenMuteTM 及 PepMuteTM Plus转染试剂，仅被发现在DMEM基础细胞培养基中偶有毒性，使用其他的培养基（含10%FBS及抗生素）培养细胞，将基本
Cell：从拓扑学角度揭示DNA复制之谜
生命分子存在缠绕的现象。但是，DNA双螺旋中那两条熟悉的链是如何在没有缠绕的情况下成功复制的，这就很难解释了。在一项新的研究中，来自美国康奈尔大学的研究人员从拓扑学角度解决了这个问题。他们研究了这种双螺旋形状对DNA复制的影响。通过使用真核生物作为模型系统，他们发现染色质（由DNA、组蛋白和非组
Cell：动态DNA或能帮助有效抵御机体基因损伤
近日，发表在国际杂志Cell上的一篇研究报告中，来自爱丁堡大学的研究人员通过研究鉴别出了DNA保护性结构的特性，这或许能够帮助研究人员对人类基因组深入理解，参与DNA支持性构架的分子往往会通过动态学变化和效应改变来应对突变；研究人员认为，这项研究或能帮助他们理解DNA损伤和基因组组织的机制，当然
Cell：科学家发现DNA修复的关键酶
日前，一项刊登在国际杂志Cell上的研究报告中，来自澳大利亚国立大学和德国海德堡大学的研究人员通过研究发现了一种DNA修复过程中的必要组分，该研究或为后期开发新型抗癌药物提供一定的思路。 研究者Tamas Fischer教授指出，当DNA被损伤后，由DNA和RNA组成的混合结构在修复遗传信息上