ExpressionLibraryScreening(Procaryotic)UsingAPFusionProteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after entering the bacterial host cell. A cDNA library is a representation of mRNA's that are expressed in a certain tissue at a certain time (e.g. rat d9 pseudopregnant decidua). Each phage contains one cDNA that was 'grabbed' by the poly-A tail and cloned into a ......阅读全文
Expression－Library－Screening－(Procaryotic)－Using－APFusion－Proteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after e
cDNA－LIBRARY－SCREENING
PREPARE SOLUTIONS1. 10mM MgSO4, 0.2% Maltose LB (100 mL):Mix 1.0 g of Bacto-Tryptone, 1.0 g of NaCl, 0.5 g of Yeast Extract, and 1.0 mL of 1M MgSO4. A
Screening－a－cDNA－Library
Screening a cDNA Libraryfor use with HybriZAP zebrafish cDNA librariesObjectivecDNA library screening allows detection of expressed genes for subseque
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 overnigh
Expression－L19－using－Pichia－pastoris
Pichia pastoris is a methylotropic yeast used to express high amounts of protein. Secreted expression is achieved with a cleavalable faktor. To yield
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
CDNA文库
CDNA文库（主要内容如下）· Construction of cDNA Library· Construction of Genome DNA Library· Library Screening OthersConstruction of cD
亲和层析实验技术方法
INTRODUCTIONThis protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of im
利用人工组合转录因子对人类基因组扫描2
Figure 5: Regulation of CDH5 by TFZFs in several human cancer cell lines.Blue, cells infected with a pMX construct containing the DNA binding domain o
蛋白质相互作用
Interaction Trap/Trap Two-Hybrid System· Yeast Two-Hybrid System (Finley Lab)This is one of the most comprehensive and detailed guide to yeast
Analysis－of－Proteins－using－Small－Format－2D－Gel－Electrophoresis
Preparation of protein samplesIntracellular virus proteinsThe following method has been developed principally for the analysis of intracellular protei
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.
人工转录因子的部件——人类锌指结构2
Table 2: Binding sites and identity of ZFPs used in VEGF activationWe then generated artificial transcription factors by fusing the three-finger domai
Twohybrid－analysis－of－genetic－regulatory－networks2
2.2 Interaction mating - large scaleWith a few modifications, the procedure described above can be used to test for interactions between a single prey
Performing－a－hunt－by－interaction－mating
AbstractWhen more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt
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
人工转录因子的部件——人类锌指结构1
Human zinc fingers as building blocks in the construction of artificial transcription factorsKwang-Hee Bae1, 4, Young Do Kwon1, 2, 4, Hyun-Chul Shin1,
cDNA
· cDNA Synthesis (Crawford Lab)mRNA can be converted into DNA (copy DNA, cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on
酵母双杂交系统
· Yeast Two-Hybrid System (Finley Lab)This is one of the most comprehensive and detailed guide to yeast two-hybrid system technique with intro
如何借助epMotion－5073l移液工作站完成微量体积的...（二）
Figure 1: epMotion 5073l worktable for qPCR setup Results and DiscussionThe automation of the qPCR setup using the 10 µL dispensing tool and the epM
Experimental－Protocol－for－cDNA－Library－Construction
Experimental Protocol for cDNA Library ConstructionIdentify appropriate celltype over-expressing corresponding gene.Find out if transcription can be s
酵母人工染色体
· Easy YAC Preparation Method (Andrew Davies,Shaw lab)· Screening YAC libraries (Donis Keller Lab)This is a method for screening YAC l
细菌人工染色体
The Construction of Bacterial Artificial Chromosome (BAC) Libraries (complete manuscript) (Clemson University Genomics Institute) Construction of BAC
Library－cDNA－Synthesis
Library cDNA Synthesis1° cDNA SynthesisN.B: During 1° cDNA synthesis, all steps should be carried while wearing gloves and all solutions should either
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
Twohybrid－analysis－of－genetic－regulatory－networks
1. Introduction and BackgroundThere is a great need for general methods to characterize the proteins that contemporary biology makes available. The li
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
高通量筛选：化合物库在药物研发过程中的应用MedChe...2
二、化合物库使用方法及使用注意事项 化合物库筛选过程主要包括化合物库选择、实验方案设计、实验进行、实验结果检测、实验方案优化等步骤。进行化合物库筛选，要根据自己的实验目的选择合适的化合物库，设计适合的实验方案，并设计好化合物作用结果的检测指标及检测方法。化合物库筛选的实验方案设计至少需要包括以下
Proteome－and－Phosphoproteome－Quantification－by－DIA－on－Orbitrap－（二）
FIGURE 2. Comparison of DDA and DIA raw data. (A) Comparison of MS1 base peak chromatograms. (B) Comparison of zoomedin chromatograms. (C) Composi
运用肽库筛选磷酸化激酶motif方法
Screening Kinase Phosphorylation Motifs Using Peptide LibrariesIsaac A. Manke and Michael B. YaffeCenter for Cancer Research, Massachusetts Institute