遗传学实验技术

This chapter describes how to create a multiplexed miRNA profiling assay using readily available technology and reagents for midplex miRNA profiles (midplex used to denote mutliplexing up to about 100 miRNAs per well). The assay described is a direct hybridization assay, i.e., it involves no ...

Microarrays have been used extensively for messenger RNA expression monitoring. Recently, microarrays have been designed to interrogate expression levels of noncoding RNAs. Here, we describe methods for RNA labeling and the use of a miRNA array to identify and measure microRNA present ...

We have developed a highly sensitive, specific, and reproducible method for microRNA (miRNA) expression profiling, using BeadArray technology. This method incorporates an enzyme-assisted specificity step, a solid-phase primer extension to distinguish between members of mi ...

MicroRNA (miRNA) profiling is of great interest because of the significant roles these short noncoding RNA molecules play in cellular regulation. Signature profiles, usually involving several miRNAs, have also been associated with dysfunctional cellular regulation such as in ca ...

In situ hybridization (ISH) is a technology that allows detection of specific nucleic acid sequences in tissue samples at the cellular level. For detection of individual microRNAs (miRNAs) and mRNAs, the ISH technology determines the cellular origin of expression and provides informa ...

Reverse transcription PCR (RT-PCR) is one of the most important techniques for analyzing RNA abundance. MicroRNAs (miRNAs) are a group of 20- to 24-nucleotide regulatory small RNAs which play an important role in plants and animals. However, the small size of miRNAs makes them difficult to be dete ...

Quantification of the microRNAs (miRNAs) in cells or tissues is a crucial step in understanding their biological functions. Development of the stem-loop reverse transcription procedure and TaqMan� miRNA assays enables accurate detection of miRNA expression levels by quantitat ...

MicroRNA analysis has been widely adopted for basic and applied science. The tools and technologies available for quantifying and analysing miRNAs are still maturing. Here, we give an introductory overview of the main tools and the challenges in their use. We also discuss the importance of bas ...

This chapter describes several methods for the isolation of miRNAs from peripheral whole blood samples or constituent fractions thereof, such as peripheral blood mononuclear cells, plasma, and serum. The methods described here are recently introduced protocols dedicated to the is ...

MicroRNAs (miRNAs) are small (∼18–25 nucleotides), endogenous, noncoding RNAs that regulate gene expression in a sequence-specific manner via the degradation of target mRNAs or the inhibition of protein translation. miRNAs are predicted to target up to one-third of all human mRNAs. Each m ...

The accurate prediction of microRNA targets is essential to understanding their function. Commonly used software produces a prohibitive number of predicted targets for each microRNA. Here, we describe procedures that refine these predictions by integrating available softwa ...

Our understanding of the importance of noncoding RNA molecules is steadily growing. One such important class of RNA molecules are microRNAs (miRNAs). These tiny RNAs fulfill important functions in cellular behavior by influencing the protein output levels of a high variety of genes thro ...

MicroRNAs (miRNAs) are short, noncoding RNAs that have the capacity to bind, capture, and silence hundreds of genes within and across diverse signaling pathways 1(Bartel, Cell 136:215–33, 2009) Specific sets of miRNAs characterize specific cell lineages of normal organisms and an increa ...

MicroRNAs (miRNAs) are an important class of gene regulators, acting on several aspects of cellular function such as differentiation, cell cycle control, and stemness. These master regulators constitute an invaluable source of biomarkers, and several miRNA signatures correlati ...

The association of dysregulated microRNAs (miRNAs) and diseases has been shown in a variety of studies. Here, we review a resource denoted as PhenomiR, providing systematic and comprehensive access to such studies. It allows machine-readable access to miRNA and target relations from the ...

Recent advances in high-throughput deep-sequencing technology have produced large numbers of short and long RNA sequences and enabled the detection and profiling of known and novel microRNAs (miRNAs) and other noncoding RNAs (ncRNAs) at unprecedented sensitivity and depth. In this ...

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Digital gene expression (DGE) profiling techniques are playing an eminent role in the detection, localization, and differential expression quantification of many small RNA species, including microRNAs (1–3). Procedures in small RNA library preparation techniques typically ...

MicroRNAs (miRNAs) are a class of small RNAs (sRNAs) of approximately 22 nucleotides in length that control eukaryotic gene expression at the translational level. They regulate a wide variety of biological processes, namely developmental timing, cell differentiation, cell prolif ...

Direct sequencing of RNA molecules using next-generation sequencing (NGS) technology has revolutionized the analysis of transcriptome with its massively parallel throughput and low cost. Here, we describe Illumina’s microRNA-Seq, a method for sequencing microRNA using the Il ...