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细胞功能测定

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Molecular Genotyping of Microbes by Multilocus PCR and Mass Spectrometry: A New Tool for Hospital Infection Control and Public Health Surveillance

We describe a new technology for the molecular genotyping of microbes using a platform known commercially as the Ibis T5000. The technology couples multilocus polymerase chain reaction (PCR) to electrospray ionization/mass spectrometry (PCR/ESI-MS) and was developed to provide r ...

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Pulsed-Field Gel Electrophoresis for Molecular Epidemiology of Food Pathogens

Foodborne diseases due to well-recognized pathogens have emerged as an important and growing public health problem with a significant impact on health. Molecular methods for subtyping these microorganisms have become a valid adjunct to the traditional techniques employed in most l ...

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Amplified Fragment Length Polymorphism Analysis

Amplified fragment length polymorphism (AFLP) analysis is a universal polymerase chain reaction (PCR)-based DNA fingerprinting technique comprising three main stages: (i) digestion of genomic DNA with restriction endonucleases and ligation to double-stranded adaptors ( ...

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Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) for the Genotyping of Bacterial Pathogens

Clustered regularly interspaced short palindromic repeats (CRISPRs) are DNA sequences composed of a succession of repeats (23- to 47-bp long) separated by unique sequences called spacers. Polymorphism can be observed in different strains of a species and may be used for genotyping. We des ...

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Sequencing of Viral Genes

The use of molecular techniques in epidemiology gives a better understanding of viral transmission and diversity, and helps to define and characterizc outbreaks. By elucidating transmission patterns and defining outbreak parameters, appropriate preventive measures can be i ...

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spa Typing for Epidemiological Surveillance of Staphylococcus aureus

The spa typing method is based on sequencing of the polymorphic X region of the protein A gene (spa), present in all strains of Staphylococcus aureus. The X region is constituted of a variable number of 24-bp repeats flanked by well-conserved regions. This single-locus sequence-based typing meth ...

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Full Sequencing of Viral Genomes: Practical Strategies Used for the Amplification and Characterization of Foot-and-Mouth Disease Virus

Nucleic acid sequencing is now commonplace in most research and diagnostic virology laboratories. The data generated can be used to compare novel strains with other viruses and allow the genetic basis of important phenotypic characteristics, such as antigenic determinants, to be elu ...

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Bacterial Genome Sequencing

For over 30 yr, the Sanger method has been the standard for DNA sequencing. Instruments have been developed and improved over time to increase throughput, but they always relied on the same technology. Today, we are facing a revolution in DNA sequencing with many drastically different platforms ...

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Genotyping of Mycobacterium tuberculosis Clinical Isolates Using IS6110-Based Restriction Fragment Length Polymorphism Analysis

A number of phylogenetic studies of Mycobacterium tuberculosis have suggested a highly clonal population structure. Despite the extreme homogeneity of M. tuberculosis strains, the genome is punctuated by a number of polymorphic regions that give rise to sufficient diversity, thus f ...

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Comparison of Molecular Typing Methods Applied to Clostridium difficile

Since the 1980s the epidemiology of Clostridium difficile infection (CDI) has been investigated by the application of many different typing or fingerprinting methods. To study the epidemiology of CDI, a typing method with a high discriminatory power, typeability, and reproducibil ...

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Multiple Locus Variable Number of Tandem Repeats Analysis

Genotyping of bacteria through typing of loci containing a variable number of tandem repeats (VNTR) might become the gold standard for many pathogens. The development of genome sequencing has shown that such sequences were present in every species analyzed, and that polymorphism exists ...

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Multilocus Sequence Typing

Multilocus sequence typing (MLST) was first proposed in 1998 as a typing approach that enables the unambiguous characterization of bacterial isolates in a standardized, reproducible, and portable manner using the human pathogen Neisseria meningitidis as the exemplar organism. S ...

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Spoligotyping for Molecular Epidemiology of the Mycobacterium tuberculosis Complex

Spacer oligonucleotide typing, or spoligotyping, is a rapid, polymerase chain reaction (PCR)-based method for genotyping strains of the Mycobacterium tuberculosis complex (MTB). Spoligotyping data can be represented in absolute terms (digitally), and the results can be readily ...

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Preparation and Crystallization of RiboswitchLigand Complexes

Riboswitches are mRNA regions that regulate the expression of genes in response to various cellular metabolites. These RNA sequences, typically situated in the untranslated regions of mRNAs, possess complex structures that dictate highly specific binding to certain ligands, such ...

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Monitoring RNALigand Interactions Using Isothermal Titration Calorimetry

Isothermal titration calorimetry (ITC) is a biophysical technique that measures the heat evolved or absorbed during a reaction to report the enthalpy, entropy, stoichiometry of binding, and equilibrium association constant. A significant advantage of ITC over other methods is that ...

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Methods for Analysis of Ligand-Induced RNA Conformational Changes

Encoded within many RNA sequences is the requisite information for folding of intricate three-dimensional structures. Moreover, many noncoding RNAs can adopt structurally distinct and functionally specialized conformations in response to specific cellular signals. The ...

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Molecular Basis of RNA-Mediated Gene Regulation on the Adenine Riboswitch by Single-Molecule Approaches

The adenine-specific pbuE riboswitch undergoes metal ion-dependent folding that involves a long-range tertiary loop–loop interaction between two stem loops. Fluorescence resonance energy transfer (FRET) and single-molecule FRET studies demonstrate the ability of the lo ...

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Umbilical Cord Blood Cells

The umbilical cord of a healthy neonate contains within it a multipotential treatment for a myriad of diseases and injuries. What was once tossed into the biohazard waste without a second thought is now known to be a goldmine of antigenically immature cells that rival the use of bone marrow for recons ...

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Human Mesenchymal Stem Cell Culture for Neural Transplantation

Mesenchymal stem cells (MSCs) have the potential to play a role in autologous repair of central nervous system injury or disease, circumventing both the complications associated with immune rejection of allogenic cells, and many of the ethical concerns associated with embryonic stem c ...

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Isolation, Expansion, and Differentiation of Adult Mammalian Neural Stem and Progenitor Cells Using the Neurosphere Assay

During development and continuing into adulthood, stem cells function as a reservoir of undifferentiated cell types, whose role is to support cell genesis in several tissues and organs. In the adult, they play an essential homeostatic role by replacing differentiated cells that are lost due ...

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