遗传学实验技术

The field of metabolomics has become increasingly important in the context of functional genomics. Together with other ”omics“ data, the investigation of the metabolome is an essential part of systems biology. Beside the analysis of human and animal biofluids, the investigation of the mi ...

NMR-based metabolomics is an analytical platform, which has been used to classify and analyze Cannabis sativa L. cell suspension cultures and plants. Diverse groups of primary and secondary metabolites were identified by comparing NMR data with reference compounds and/or by structu ...

In many RNA silencing applications, there is a benefit to expressing multiple interfering RNAs simultaneously. This can be achieved by using a single RNA polymerase II promoter to express multiple micro(mi)RNA-formatted interfering RNAs that are arranged in a polycistronic cluster, ...

Antisense oligodeoxyribonucleotides have been used for decades to achieve sequence-specific silencing of gene expression. However, all early generation oligonucleotides (e.g., those with no other modifications than the phosphorothioate backbone) are inactive in vitro u ...

In order to unfold the function of genes, solely performing mRNA over-expression is not enough nowadays. Traditional protein expression experiments, such as Western blotting and immunohistochemical staining, could only provide researchers the changes of expression levels and ...

We have developed four expression vectors to express antisense RNAs (asRNAs) by which genes of interest are silenced in Escherichia coli. The vectors are all IPTG-inducible and co-transformable in any combination and target genes are silenced conditionally and concurrently. Furth ...

RNA interference (RNAi) enables the suppression, and hence the functional analysis, of individual genes. The use of the tetracycline (tet)-controlled transcription activation system for RNAi has become a valuable tool for conditional gene inactivation both in vitro and in vivo. Here, t ...

Matrix-based yeast two-hybrid screens are an alternative to library-based screens. Recent improvements of matrix screens (also called array screens), use various pooling strategies as well as novel vectors that increase its efficiency while decreasing the false-negative rate, t ...

Analyzing the putative interaction partners of an individual protein is one approach to elucidate its function. In the LuMPIS protocol, bait and prey proteins are expressed with N-terminal maltose binding protein (MBP)- and eGFP-luciferase (eGFP-luc) tags, respectively. Positive p ...

In the field of proteomics, numerous advanced technologies have evolved that aim to provide the molecular data necessary for an in-depth understanding of biological processes. Protein–protein interactions (PPI) are at the heart of cellular function and a milestone yet to be achieved is t ...

Microscale thermophoresis (MST) is a new method that enables the quantitative analysis of molecular interactions in solution at the microliter scale. The technique is based on the thermophoresis of molecules, which provides information about molecule size, charge, and hydration s ...

We have developed a rapid, high-throughput assay for measuring the catalytic activity (DNA supercoiling or relaxation) of DNA topoisomerases. The assay utilizes intermolecular triplex formation between an immobilized triplex-forming oligo (TFO) and a triplex-forming regi ...

Major technological advances have made proteomics an extremely active field for biomarker discovery in recent years due primarily to the development of newer mass spectrometric technologies and the explosion in genomic and protein bioinformatics. This leads to an increased emph ...

Proteolysis is the key step for proteomic studies integrated with MS analysis. Compared with the conventional method of in-solution digestion, proteolysis by a protease-immobilized microreactor has a number of advantages for proteomic analysis; i.e., rapid and efficient digesti ...

Technological advances in the field of microarray production and analysis lead to the development of protein microarrays. Of these, antigen microarrays are one particular format that allows the study of antigen–antibody interactions in a miniaturized and highly multiplexed fas ...

Two-dimensional gel electrophoresis (2-DE) is the most popular and versatile method of protein separation among a rapidly growing array of proteomic technologies. Based on two independent biochemical characteristics of proteins, it combines isoelectric focusing, which sepa ...

The ability to profile bacterial gene expression has markedly advanced the capacity to understand the molecular mechanisms of pathogenesis, epidemiology, and therapeutics. This advance has been coupled with the development of techniques that enable investigators to identify ...

RNA editing can lead to amino acid substitutions in protein sequences, alternative pre-mRNA splicing, and changes in gene expression levels. The exact in vivo modes of interaction of the RNA editing enzymes with their targets are not well understood. Alterations in RNA editing have been link ...

We present a quantification method for affinity-based DNA microarrays which is based on the real-time measurements of hybridization kinetics. This method, i.e., real-time DNA microarrays, enhances the detection dynamic range of conventional systems by being impervious to probe sa ...

Pituitary tumor transforming gene is an important gene which is involved in many cellular functions including cell division, DNA repair, organ development, expression, and secretion of various angiogenic and metastatic factors. Overexpression of this gene has also been reported in ...