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丁香实验推荐阅读
Sequencing of the Rat Genome and Databases

The rat is an important system for modeling human disease. Four years ago, the rich 150-year history of rat research was transformed by the sequencing and annotation of the rat genome, ushering in an era of exceptional opportunity for identifying genes and pathways underlying disease phenoty ...

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Inducible and Conditional Promoter Systems to Generate Transgenic Animals

Transgenic animals are very useful models that can be utilized for the analysis of temporal and spatial gene expression in vivo. However, generation of a transgenic animal may become problematic if the presence of the transgene leads to conditions which are toxic or lethal to cell growth. In an eff ...

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Generation of Transgenic Rats by Microinjection of Short DNA Fragments

Here we describe an efficient technique to generate transgenic rats by microinjection of short DNA fragments. We have focused on optimal conditions for superovulation of prepubescent females Sprague–Dawley (CD) strains to have good quality embryos, pseudopregnant females, zyg ...

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Generation of Transgenic Rats Using YAC and BAC DNA Constructs

Transgenic rats with a simple plasmid vector smaller than 20 Kb show insufficient expression and tissue specificity of the introduced transgene. Vectors derived from yeast artificial chromosome (YAC) and bacterial artificial chromosome (BAC), consisting of DNA fragments up to ∼1 Mb ( ...

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The Use of Lentiviral Vectors to Obtain Transgenic Rats

Lentiviral vectors are now well recognized as good vehicles for gene delivery. This is because they can efficiently transduce both dividing and post-mitotic cells, and stably integrate into the host genome allowing for long-term expression of the transgene. Their potential utility for ...

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Generation of Transgenic Rats by Ooplasmic Injection of Sperm Cells Exposed to Exogenous DNA

Intracytoplasmic sperm injection (ICSI) has been successfully achieved in mice and rats using a piezo-driven injection pipette, with the offspring rate of 30%. The ICSI technique was applied not only to rescue infertile male strains but also to produce transgenic rodents. The ICSI-media ...

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Methylated DNA Immunoprecipitation Genome-Wide Analysis

DNA methylation is an important and potentially heritable component of the epigenetic machinery that has a major role in the control of gene expression and can be deregulated in many diseases. This modification of genomic DNA can be assessed using the methylated DNA immunoprecipitation ( ...

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SIRPH: An HPLC-Based SNuPE for Quantitative Methylation Measurement at Specific CpG Sites

Genome-wide sequence-specific methylation analysis has become a readily available and affordable procedure in epigenetic laboratories. Most of these procedures are based on immunoprecipitation, microarrays, or next generation deep bisulfite sequencing. However, most ...

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A Combined Bisulfite Restriction Analysis Bioinformatics Tool: Methyl-Typing

In this chapter, we introduce our developed freeware tool Methyl-Typing. It provides methylation-related bioinformatics with a special focus on combined bisulfite restriction analysis (COBRA). We give an overview of the implementation and program modules for Methyl-Typing. Va ...

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Closed-Tube PCR Methods for Locus-Specific DNA Methylation Analysis

Closed-tube PCR methods (sometimes referred to as in-tube PCR methods) for locus-specific DNA �methylation analysis are methodologies in which the amplification and analysis of bisulphite-modified DNA take place in one tube without the need to remove the PCR products for further anal ...

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Analysing DNA Methylation Using Bisulphite Pyrosequencing

Bisulphite pyrosequencing is a quantitative methodology for the investigation of DNA methylation of sequences up to 100-bp in length. Biotin-labelled, single-stranded PCR products generated from bisulphite-treated DNA are used as a template with an internal primer to perform the p ...

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Genome Acquisition in Horizontal Gene Transfer: Symbiogenesis and Macromolecular Sequence Analysis

Phylogenetic diagrams (“trees of life”) based on computer-generated analyses of nucleic acid (DNA, RNA) or protein (amino acid residues) sequences are purported to reconstruct evolutionary history of the living organisms from which the macromolecules were isolated (1). “Horizon ...

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Gene Transfer: Who Benefits

Horizontal gene and genome transfer forces us to recognize that life evolves by fusion as well as bifurcation of lineages, and necessitates the expansion of traditional views of evolution. This chapter reviews the role that horizontal gene transfer (HGT) may play in integrating selection ...

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Defining the Mobilome

This chapter defines the agents that provide for the movement of genetic material which fuels the adaptive potential of life on our planet. The chapter has been structured to be broadly comprehensive, arbitrarily categorizing the mobilome into four classes: (1) transposons, (2) plasmids, ...

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The Interplay of Homologous Recombination and Horizontal Gene Transfer in Bacterial Speciation

Bacteria experience recombination in two ways. In the context of the Biological Species concept, allelic exchange purges genic variability within bacterial populations as gene exchange mediates selective sweeps. In contrast, horizontal gene transfer (HGT) increases the size of ...

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Epistemological Impacts of Horizontal Gene Transfer on Classification in Microbiology

We describe the reasons why the newly recognized process of horizontal gene transfer (HGT) forces evolutionists who study classification and microbiology to go beyond the classical Darwinian framework. We recall the importance of processes in philosophical definitions of speci ...

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Prefractionation, Enrichment, Desalting and Depleting of Low Volume and Low Abundance Proteins and Peptides<br>Using the MF10

The success of proteomics relies heavily in the ability to characterize very diverse species of proteins. This diversity stems from a proteins physicochemical properties, its copy number and abundance and its association with other proteins. Prefractionation and simplificati ...

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Microscale Isoelectric Focusing in Solution: A Method for Comprehensive and Quantitative Proteome Analysis Using 1-D and 2-D DIGE Combined with MicroS

Current methods for quantitatively comparing complex protein profiles such as two-dimensional gel electrophoresis (2-DE), 2-D differential in-gel electrophoresis (DIGE), and liquid chromatography (LC)-mass spectrometry (MS) have limited resolution and dynamic ranges ...

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Fractionation of Complex Protein Mixtures by Liquid-Phase Isoelectric Focusing

Protein fractionation is essential to uncovering low-abundance proteins in complex protein mixtures. Many common methods and techniques are used to fractionate proteins, including chromatography (size exclusion, affinity, ion exchange, etc.), electrophoresis, and solut ...

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End-Labeling of DNA Probes

End-labeling is a rapid and sensitive method for radioactively, or nonisotopically, labeling DNA fragments and is useful for visualizing small amounts of DNA. End-labeling can also be used to label fragments at one end. All of the enzymes employed are specific to either the 3′ or 5′ termini of DNA and wil ...

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