遗传学实验技术

Laser capture microdissection (LCM) has become a powerful technique that allows analyzing gene expression in specific target cells from complex tissues. It is widely used in animal research, yet few studies on plants have been carried out. We have applied this technique to the plants–nemat ...

RNA in situ hybridization using digoxigenin-labeled riboprobes on tissue sections is a powerful technique for revealing microscopic spatial gene expression. Here, we describe an in situ hybridization method commonly practiced in Arabidopsis research labs. The highly stringe ...

Over the past few years only, next-generation sequencing technologies became accessible and many applications were rapidly derived, such as the development of RNA-seq, a technique that uses deep sequencing to profile whole transcriptomes. RNA-seq has the power to discover new transc ...

Second-generation DNA sequencing platforms have emerged as powerful tools in biological research. Their high sequence output at lower cost and minimal input DNA requirement render them suitable for broad applications ranging from gene expression studies to personalized clini ...

The ribonuclease protection assay (RPA) has emerged as an important methodology for the detection, mapping, and quantification of RNAs. In this assay, total or cytoplasmic RNAs are hybridized to a high-specific activity antisense radioactive RNA probe synthesized by in vitro transcr ...

SuperSAGE is a variant of the Serial Analysis of Gene Expression (SAGE) technology, based on counting transcripts by sequencing analysis of short sequence tags. In SuperSAGE, 26 bp tags are extracted from cDNA using the Type III restriction endonuclease EcoP15I. The use of a longer tag size in Sup ...

Next-generation sequencing (NGS) is becoming a routine experimental technology. It has been a great success in recent years to profile small-RNA species using NGS. Indeed, a large quantity of small-RNA profiling data has been generated from NGS, and computational methods have been develo ...

Using High-Throughput DNA Sequencing (HTS) to examine gene expression is rapidly becoming a �viable choice and is typically referred to as RNA-seq. Often the depth and breadth of coverage of RNA-seq data can exceed what is achievable using microarrays. However, the strengths of RNA-seq are oft ...

The method for the construction of Expressed Sequence Tag (EST) assemblies described here uses reads generated from 454 pyrosequencing and Sanger and Illumina (Solexa) sequencing technologies as input. It is consistent with and parallels many established EST assembly protocols, f ...

Small RNAs (sRNAs) have emerged as one of the most important regulators of gene expression in eukaryotes. sRNAs are intermediate molecules as well as end products in the antiviral defense pathway called RNA interference in plants and animals. Profiling of sRNAs using next-generation sequ ...

Small RNAs are short noncoding RNAs with important regulatory roles in many cellular processes. Small RNAs are generated by DICER or DICER-like proteins and then incorporated into RNAi effector �proteins ARGONAUTEs (AGOs) for silencing of their targets. In plants, small RNAs regulate h ...

Due to crucial roles in gene regulation, noncoding small RNAs (smRNAs) of 20–30 nucleotides (nt) have been intensively studied in mammals and plants, and are known to be implicated in significant diseases and metabolic disorders. Elucidation of biogenesis mechanisms and functional cha ...

MicroRNAs (miRNAs) play important roles in development in plants, and some miRNAs show developmentally regulated organ- and tissue-specific expression patterns. Therefore, in situ detection of mature miRNAs is important for understanding the functions of both miRNAs and their ta ...

Northern blotting is a valuable method for detection and quantification of RNA in the field of virology. Although many methods including a various versions of polymerase chain reaction have been developed over the years, Northern blotting has been still considered as a useful and effective ...

The complexity of molecular diagnostic assays is a significant barrier to employment in point-of-care diagnostics, despite the growing need for such technologies. We have developed a sensitive, accurate, rapid, and simple DNA amplification scheme that shows potential for transla ...

A multiple primer extension (MPEX) was originally developed for the hybridization, extension, and amplification of a DNA template on a planar substrate by Kinoshita et al. in 2006. Herein we present a modified MPEX method refined by our group for single nucleotide polymorphism (SNP) detecti ...

The increasing need for large-scale genotyping applications of single nucleotide polymorphisms (SNPs) in model and nonmodel organisms requires the development of low-cost technologies accessible to minimally equipped laboratories. The method presented here allows effi ...

Progress in single nucleotide polymorphism (SNP) detection technologies has provided information for SNP-based studies, such as identification of candidate genes for the complex genetic diseases, pharmacogenetic analysis, drug development, population genetics, evol ...

A microarray-based method for detecting single nucleotide polymorphisms (SNPs) using solid-phase polymerase chain reaction (PCR) on magnetic nanoparticles (MNPs) was developed. In this method one primer with a biotin label is captured by streptavidin-coated MNPs (SA-MNPs), and t ...

Single nucleotide polymorphisms (SNPs) are the most common form of polymorphisms present in the human genome. The single base primer extension (SBE) method is an effective and sensitive tool that can type over 30 known loci scattered throughout an organism’s genome in a single reaction. It all ...