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基因表达差异显示实验

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Genome-Scale Probe and Primer Design with PRIMEGENS

This chapter introduces the software package PRIMEGENS for designing gene-specific probes and associated PCR primers on a large scale. Such design is especially useful for constructing cDNA or oligo microarray to minimize cross-hybridization. PRIMEGENS can also be used for design ...

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Fast Masking of Repeated Primer Binding Sites in Eukaryotic Genomes

In this article, we describe the working principle and a list of practical applications for GenomeMasker—a program that finds and masks all repeated DNA motifs in fully sequenced genomes. The GenomeMasker exhaustively finds and masks all repeated DNA motifs in studied genomes. The softwa ...

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Degenerate Primer Design: Theoretical Analysis and the HYDEN Program

A polymerase chain reaction (PCR) primer sequence is called degenerate if some of its positions have several possible bases. The degeneracy of the primer is the number of unique sequence combinations it contains. We study the problem of designing a pair of primers with prescribed degeneracy t ...

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SNPbox: Web-Based High-Throughput Primer Design with an Eye for Repetitive Sequences

In this omics era, researchers in biosciences need more than ever user-friendly, fast, and straightforward computational tools for high-throughput research. In this chapter, we present SNPbox and explain the general primer design strategy. We also show how the four modules, Exon, Single ...

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An Iterative Method for Selecting Degenerate Multiplex PCR Primers

Single-nucleotide polymorphism (SNP) genotyping is an important molecular genetics process, which can produce results that will be useful in the medical field. Because of inherent complexities in DNA manipulation and analysis, many different methods have been proposed for a stand ...

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Primer Design for Multiplexed Genotyping

Single-nucleotide polymorphism (SNP) genotyping can be carried out by annealing an oligonucleotide primer directly adjacent to the polymorphism and carrying out a single base extension using a polymerase reaction with labeled dideoxynucleotide triphosphates. This can be mu ...

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MultiPLX: Automatic Grouping and Evaluation of PCR Primers

In this chapter, we describe MultiPLX—a tool for automatic grouping of PCR primers for multiplexed PCR. Both generic working principle and step-by-step practical procedures with examples are presented. MultiPLX performs grouping by calculating many important interaction leve ...

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MultiPrimer: A System for Microarray PCR Primer Design

To construct full-genome spotted microarrays, a large number ofPCR primers that amplify the required DNA need to be synthesized.We describe an algorithmic technique that allows one to use fewerprimers to achieve this goal. This can reduce the expense of primers are usually designed, so that ...

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AlleleID: A Pathogen Detection and Identification System

Efficient clinical diagnosis of pathogens is important for the management of infectious diseases. Conventional methods have longer turnaround time and, in most cases, lower sensitivity. Nucleic acid-based methods for the detection of microorganisms are rapid, sensitive and are g ...

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Modified Oligonucleotides as Tools for Allele-Specific Amplification

Allele-specific polymerase chain reaction (PCR), a method that reports nucleotide variations through either the presence or the absence of a DNA product obtained through PCR amplification, holds the promise to combine target amplification and analysis in one single step. Recently, it ...

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Designing Primers for Whole Genome PCR Scanning Using the Software Package GenoFrag: A Software Package for the Design of Primers Dedicated to Whole-G

Whole-genome polymerase chain reaction (PCR) scanning (WGPS) is based on the PCR amplification of small-sized chromosomes (e.g., bacterial chromosomes) by long-range PCR with a set of primers designed using a reference strain and applied to amplify several other strains. Such an approach ...

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Designing PCR Primer for DNA Methylation Mapping

DNA methylation is an epigenetic mechanism of gene regulation, and aberrant methylation has been associated with various types of diseases, especially cancers. Detection of DNA methylation has thus become an important approach for studying gene regulation and has potential diagn ...

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BiSearch: ePCR Tool for Native or Bisulfite-Treated Genomic Template

The design of adequate primers for polymerase chain reaction (PCR) is sometimes a difficult task. This is the case when either the target sequence harbors unusual nucleotide motifs or the template is complex. Unusual nucleotide motifs can be repeat elements, whereas complex templates are t ...

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Graphical Design of Primers with PerlPrimer

PerlPrimer is a cross-platform application for the design of standard PCR, bisulfite PCR, real-time PCR, and sequencing primers. The program combines accurate primer-design algorithms with powerful interfaces to commonly used Internet databases, such as sequence retrieval from ...

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Identifying Nonspecific SAGE Tags by Context of Gene Expression

Many serial analysis of gene expression (SAGE) tags can be matched to multiple genes, leading to difficulty in SAGE data interpretation and analysis. As only a subset of genes in the human genome are transcribed in a certain type of tissue/cell, we used microarray expression data from different ti ...

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Clustering Analysis of SAGE Transcription Profiles Using a Poisson Approach

To gain insights into the biological function and relevance of genes using serial analysis of gene expression (SAGE) transcription profiles, one essential method is to perform clustering analysis on genes. A successful clustering analysis depends on the use of effective distance or si ...

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Scaling of Gene Expression Data Allowing the Comparison of Different Gene Expression Platforms

Serial analysis of gene expression (SAGE) and microarrays have found a widespread application, but much ambiguity exists regarding the amalgamation of the data resulting from these technologies. Cross-platform utilization of gene expression data from the SAGE and microarray tec ...

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Correction of Technology-Related Artifacts in Serial Analysis of Gene Expression

Serial analysis of gene expression (SAGE) is a powerful technique for measuring global gene expression through sampling of transcript tags. SAGE tag collections or libraries serve as a rich data source for differential gene expression analysis, transcriptome mapping, and gene disco ...

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Alu-PCR Fingerprinting of YACs

Random fingerprinting strategies have been applied to a number of genome projects using unordered collections of phage or cosmid clones, e.g., Escherichia coli (1), Saccharomyces cerevisiae (2), and Caenorhabditis elegans (3). Overlaps were identified between either phage or cosmid ...

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Detection of Chimerism in YAC Clones

Chimerism, the presence of noncontiguous DNA fragments in the same clone, is one of the most common problems encountered when working with yeast artificial chromosomes (YACs). Its frequency can vary among the different libraries, but on average, 40–60% of YAC clones among the most used librari ...

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