遗传学实验技术

RNA silencing is a sequence-specific RNA degradation mechanism conserved in eukaryotes including fungi, plants, and animals. One of the three RNA silencing pathways is DNA methylation which is the result of interaction between DNA and siRNA, a hallmark of RNA silencing. Bisulfite sequen ...

MicroRNAs (miRNAs) are a specific class of 21-nt small RNAs. They regulate the expression of specific target genes by various types of post-transcriptional regulation mechanisms, such as transcript cleavage and translation suppression. The biological function of an miRNA is theref ...

Deep sequencing technologies have become very powerful tools in the identification and quantification of small RNAs involved in gene regulation. Small interfering RNA (siRNA) and miRNA are two classes of DCL-dependent small RNAs known to affect phenotype, developmental regulati ...

Plant microRNAs (miRNAs) are a class of endogenous small RNAs that are essential for plant development and survival. They arise from larger precursor RNAs with a characteristic hairpin structure and regulate gene activity by targeting mRNA transcripts for cleavage or translational r ...

Hybridization of labeled cDNA to microarrays is an intuitively simple and a vastly underestimated process. If it is not performed, optimized, and standardized with the same attention to detail as e.g., RNA amplification, information may be overlooked or even lost. Careful balancing of the am ...

Much information can be obtained from knowledge of the relative expression level of each gene in the transcriptome. With the current advances in technology as little as a single cell is required as starting material for gene expression experiments. The mRNA from a single cell may be linearly ampl ...

Many steps of optimization are needed to achieve large-scale fabrication of high-quality DNA microarrays. These steps involve the printing instrument, the probes to be printed, microarray slides, and spotting buffer together with the surrounding environment, such as humidity and t ...

Among the parameters which influence the success of a microarray experiment, the attachment of the nucleic acid captures to the support surface plays a decisive role. This article attempts to review the main concepts and ideas of the multiple variants which exist in terms of the immobilization ...

There are many ways to fabricate DNA microarrays. The four main types of arrays are spotted arrays of pre-made DNA probes, in situ synthesis of DNA arrays, random bead arrays and suspension arrays. The different types of array can address different biological assays. Spotted arrays are suitable f ...

Whole genome association studies have recently been enabled by combining tag SNP information derived from the International HapMap project with novel whole genome genotyping array technologies. In particular, Infinium� whole genome genotyping (WGG) technology now has the power ...

Microarray-based Comparative Genomic Hybridization (array-CGH) has been applied for a decade to screen for submicroscopic DNA gains and losses in tumor and constitutional DNA samples. This method has become increasingly flexible with the integration of new biological resources ...

Since all measurements from a DNA microarray is dependant on the probes used, a good choice of probes is of vital importance when designing custom microarrays. This chapter describes how to design expression arrays using the OligoWiz software suite. The desired general features of good prob ...

DNA microarrays can be used for large number of application where high-throughput is needed. The ability to probe a sample for hundred to million different molecules at once has made DNA microarray one of the fastest growing techniques since its introduction about 15 years ago. Microarray tec ...

Chromatin immunoprecipitation (ChIP) is a powerful procedure to investigate the interactions between proteins and DNA. ChIP-chip combines chromatin immunoprecipitation and DNA microarray analysis to identify protein–DNA interactions that occur in vivo. This genome-wi ...

Array-CGH involves the comparison of a test to a reference genome using a microarray composed of target sequences with known chromosomal coordinates. The test and reference DNA samples are used as templates to generate two probe DNAs labeled with distinct fluorescent dyes. The two probe DNAs ...

The spatial resolution of microarray-based comparative genomic hybridization (array-CGH) is dependent on the length and density of target DNA sequences covering the chromosomal region of interest. Here we describe the methods developed at the Wellcome Trust Sanger Institute (Ca ...

Microarray technology has revolutionized the detection and analysis of microbial pathogens. The success of this technology is evident from the various microarrays that have been developed for this purpose, variation in the density of probes, and the time ranges required for assay comp ...

The need for multiplexed methods for SNP genotyping has rapidly increased during the last decade. We present here a flexible system that combines highly specific genotyping by minisequencing single-base extension with the advantages of a microarray format that allows highly multip ...

In this review, we describe the laboratory implementation of Infinium� whole genome genotyping (WGG) technology for whole genome association studies and copy number studies. Briefly, the Infinium WGG assay employs a single tube whole genome amplification reaction to amplify the ent ...

In any microarray hybridization experiment, there are contributions at each probe spot due to the match and numerous mismatch target species (i.e., cross-hybridizations). One goal of temperature optimization is to minimize the contribution of mismatch species; however, achieving ...