遗传学实验技术

Two-dimensional (2D) gel electrophoresis employs distinct electrophoretic conditions to better resolve complex mixtures of molecules. In combination with Southern analysis, 2D agarose gel electrophoresis is routinely employed to detect and analyze DNA intermediates th ...

The analysis of replication intermediates by the neutral–neutral two-dimensional agarose gel technique allows determining the chromosomal positions where DNA replication initiates, whether replication forks pause or stall at specific sites, or whether two DNA molecules un ...

The neutral–neutral two-dimensional agarose gel technique is mainly used to determine the chromosomal positions where DNA replication starts, but it is also applied to visualize replication fork progression and breakage as well as intermediates in DNA recombination. Here we provi ...

Branched DNA molecules are generated by the essential processes of replication and recombination. Owing to their distinctive extended shapes, these intermediates migrate differently from linear double-stranded DNA under certain electrophoretic conditions. However, t ...

Presented here is a complete molecular cloning protocol consisting of a number of separate but interconnected methods such as preparation of E. coli competent cells; in vitro DNA digestion and ligation; PCR; DNA agarose gel electrophoresis and gel extraction; and screening transforma ...

The technique of gel electrophoresis is now firmly established as a routine laboratory method for analyzing DNA. Here, we describe the development of the methodology as well as a brief explanation of how the technique works. There is a short introduction to pulsed-field agarose gel electroph ...

Telomerase is the enzyme that extends the chromosome ends, thereby contributing to eukaryotic cell genome stability. Telomerase is expressed in the majority of cells that have an unlimited proliferation such as stem cells and cancer cells. The increased interest in telomerase in cancer ...

Protein-induced DNA bending plays a key role in many essential biological processes, such as DNA replication, recombination, and transcription, and can be analyzed by a variety of biochemical and biophysical methods, such as electrophoresis mobility shift assay (EMSA), X-ray crysta ...

DNA-binding proteins play essential roles in many cellular processes. Understanding on a molecular level how these proteins interact with their cognate sequences can provide important functional insights. Here, we describe a band shift assay in agarose gel to assess the mode of protein b ...

The band-shift assay using polyacrylamide gel electrophoresis is a powerful technique used to investigate DNA–protein interactions. The basis of the method is the separation of free DNA from DNA–protein complexes by virtue of differences in charge, size, and shape. The band-shift assay c ...

Fluorescence in situ hybridization (FISH) to label fragments of DNA with probes which can specifically locate a genomic region of interest, combined with the single cell electrophoresis (Comet) assay, also termed Comet-FISH, allows the quantification of DNA damage and repair at a specif ...

The comet assay or single-cell gel electrophoresis assay is a relatively simple and sensitive technique for quantitatively measuring DNA damage and repair at the single-cell level in all types of tissue where a single-cell suspension can be obtained. Isolated cells are mixed with agarose, ...

Pulsed-field gel electrophoresis (PFGE) is a technique that resolves chromosome-sized DNA molecules in an agarose gel. As well as DNA mapping and karyotyping applications, PFGE techniques are well adapted to follow DNA rearrangements over time in a quantitative manner. Because of the ve ...

The separation of fragments of DNA by agarose gel electrophoresis is integral to laboratory life. Nevertheless, standard agarose gel electrophoresis cannot resolve fragments bigger than 50 kb. Pulsed-field gel electrophoresis is a technique that has been developed to overcome the l ...

Denaturing urea polyacrylamide gel electrophoresis (PAGE) allows the separation of linear single-stranded DNA molecules based on molecular weight. This method can be used to analyze or purify short synthesized DNA oligonucleotides or products from enzymatic reactions. In this c ...

Temperature Gradient Gel Electrophoresis (TGGE) is a form of electrophoresis in which temperature gradient is used to denature molecules as they move through either acrylamide or agarose gel. TGGE can be applied to analyze DNA, RNA, protein–DNA complexes, and, less commonly, proteins. Se ...

Two main hurdles hinder the widespread acceptance of plants as a preferred protein expression platform: low accumulation levels and expensive chromatographic purification methods. Fusion of proteins of interest to fungal hydrophobins has provided a tool to address both accumul ...

All proteins fold into a defined three-dimensional shape that is compatible with the cellular role and/or biological activity of those proteins. Molecular chaperones are a family of proteins whose role is to assist the folding and targeting of proteins in both normal and stressed cells. The ra ...

Historically, it has been proved difficult to adapt the traditional baculovirus expression systems to an automated platform because of the complexity of the processes involved. One of the major bottlenecks is the selection of recombinant from parental viruses. We have developed a bacm ...

Plant-based expression technologies for recombinant proteins have begun to receive acceptance for pharmaceuticals and other commercial markets. Protein products derived from plants offer safer, more cost-effective, and less capital-intensive alternatives to traditi ...