ConstructionandManipulationofLargeInsertBacterialCloneLibraries
Acknowledgements The organizer of the workshop acknowledges Dr. Murray Milford, Professor and Interim Head, and Dr. Mark Hussey, Professor and Interim Associate Head, Department of Soil and Crop Sciences, Texas A&M University for their valuable suggestions and support, and Ms. Chantel Scheuring and Ms. Elizabath Huff for their assistance in organizing the workshop. The author of the manual thanks Ms. Chantel......阅读全文
Microaspiration－of－Esophageal－Gland－Cells－and－cDNA－Library－Construction－for
Microaspiration of Esophageal Gland Cells and cDNA Library Construction for Identifying Parasitism Genes of Plant-Parasitic NematodesIdentifying p
基于epMotion－5075t系统与KPPA－HyperPlus试剂盒的全自动测序..1
基于epMotion 5075t系统与KPPA HyperPlus试剂盒的全自动测序前文库制备方案Automated KAPA HyperPlus DNA Library Preparation for Illumina® Sequencing on the Eppendorf epMotion®
Hydrolytic－Activity－of－Bacterial－Supernatants－for－Fungal－Suppression
As the fungal growth suppression by biocontrol agents (BCA) in solid media using dual plate assay has some issues regarding nutrient limitation. A pr
Bacterial－Expression－of－GSTfusion－Proteins
1. Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2. Grow larger culture (100x volume of starter culture) using the ove
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates
Abstract The kinetic characteristics and regulation of aspartate carbamoyltransferase activity were studied in lysates and cell extracts of He
TritonPrep－Method－for－bacterial－DNA－Purification
Triton-Prep Method for bacterial DNA PurificationGrow 5 ( large scale-15ml culture). Harvest in single eppendorf tube (or 15 ml disposable tube).Resus
细菌转化（bacterial－transformation）原理和操作
1．目的学会质粒DNA转化感受态受体菌的技术。2．原理质粒DNA粘附在细菌细胞表面，经过42°C短时间的热击处理，促进吸收DNA。然后在非选择培养基中培养一代，待质粒上所带的抗菌素基因表达，就可以在含抗菌素的培养基中生长。3．器材旋涡混合器，微量移液取样器，移液器吸头，1.5ml 微量离心管，双面微
利用人工组合转录因子对人类基因组扫描
Scanning the human genome with combinatorial transcription factor librariesPublished online: 18 February 2003, doi:10.1038/nbt794March 2003 Volume 21
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
DNA的酶学操作
DNA的酶学操作DNA Modifying Enzymes (Michael Blaber)Introduction to bacterial restriction/modification system. It provides very useful background knowledge
基于epMotion－5075t系统与KPPA－HyperPlus试剂盒的全自动测序...2
Results and DiscussionThe post-ligation qPCR results were used to calculate the percentage of starting material that was successfully adapter ligate
Preparation－of－Bacterial－Proteins－for－Analysis－by－2DPAGE
The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was prin
基因克隆(gene－clone)的几种常用方法介绍1
基因(gene)是遗传物质的最基本单位,也是所有生命活动的基础。不论要揭示某个基因的功能,还是要改变某个基因的功能,都必须首先将所要研究的基因克隆出来。特定基因的克隆是整个基因工程或分子生物学的起点。本文就基因克隆的几种常用方法介绍如下。1 根据已知序列克隆基因对已知序列的基因克隆是基因克隆方法中最
基因克隆(gene－clone)的几种常用方法介绍2
其基本原理是：用一个在种源上相近的基因组将靶基因组中所有共同的基因掩盖起来,而只暴露出特异的基因,在整个反应中只有特异基因能被扩增。其操作程序为:(1)用同一限制性内切酶(一般用Bam HI,Bgl II或Hind III)同时处理靶基因组和掩盖基因组DNA，其中掩盖基因组DNA的量至少要2
Packagene®－Lambda－DNA－Packaging－System
Packagene® Lambda DNA Packaging SystemThe Packagene® Lambda DNA Packaging System is derived from the unique one-strain host system (Rosenberg). The Sy
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
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates3
Different enzyme assays for ACTase study in H. pylori ACTase properties were studied in situ in cell-free extracts to obtain information on enzyme f
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates4
The use of a micro-titre based colorimetric assay provided a third method for the study of ACTase activity, and the most useful for large scale measu
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates2
Measurement of ACTase activityNuclear magnetic resonance spect roscopy (NMR)The unique potential of NMR spectroscopy for monitoring simultaneously the
Screening－Bacterial－Colonies－Using－Xgal－and－IPTG:－αComplementation
实验概要 α-complementation occurs when two inactive fragments of E. coli β-galactosidase associate to form a functional enzyme. Many plasmid
454高通量测序——研究土壤微生物的新手段（三）
注：已用454焦磷酸测序法发表的文章总结1、Archaea predominate among ammonia-oxidizing prokaryotes in soils—— (2006) Nature 442: 806-809——if 36.1012、454 Pyrosequencing ana
Bacterial－Expression－of－IRS1－containing－GSTfusion－Proteins
1. Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2. Grow larger culture using the overnight culture as a seeding culture.
重组DNA的分离、克隆与测序实验手册3
G. Bacterial cell maintenanceFour strains of E. coli are used in these studies: JM101 for M13 infection and isolation (4), XL1BMRF' (Stratagene) f
基因可隆的方法
Serial Analysis of Gene Expression (SAGE) SAGE is a powerful tool that allows the analysis of overall gene expression patterns with digital analys
Phage－Titer
IntroductionLambda phage is a commonly used vector for transgenes. Its very high rate of infectivity makes it ideal for creating large numbers of clon
How－to－build－a－BAC－library
Introduction The most important aspect of our cloning vectors is that they are based on the E. coli F-factor replicon. It allows for strict
Human－T－Cell－Activation－with－antiCD3－(clone－UCHT1－or－HIT3a)
实验概要T-cell activation, which plays a central role in the regulation of immune responses, involves multiple intracellular signaling events originatin
The－TRC－shRNA－Design－Methods－and－Rules
OverviewWe design shRNA molecules with an algorithm. Our algorithm uses several criteria to rank potential 21mer targets within each human and mouse R
Competitive－RTPCR－Strategy－for－Quantitative－Evaluation－2
Determination of Accuracy of the Competitive PCRTo test the precision of the results obtained with this competitive PCR, five different amounts of T (