遗传学实验技术

With the completion of the draft sequence of the human genome (1,2), it is now essential to extract biological information from the large volumes of human genomic sequence data. A number of attempts, which can be comprehensively termed “functional genomics,” are being carried out to decipher wh ...

The generation of a complementary DNA (cDNA) library using transcriptional amplification has been developed to offer better linear amplification than polymerase chain reaction (PCR)-based methods. By incorporating a RNA promoter element during reverse transcription of mes ...

The secondary structure of most functional RNA molecules is strongly conserved in evolution. Prediction of these conserved structures is therefore of particular interest when studying noncoding RNAs. Moreover, structure predictions on the basis of several sequences produce m ...

Gene expression technology offers great potentials to generate new insights into human disease pathogenesis; however, the data quality remains a major obstacle for realizing its potentials. In the present study 60-mers oligonucleotide target immobilized on coated glass slides w ...

Array-based comparative genomic hybridization (array CGH) is becoming a prominent genomic technology with many important applications in biomedical research. Although several platforms of this technology have been published, successful implementation of this technol ...

Optimization of oligonucleotide DNA microarrays is a multiparametric problem. The goal of the optimization process is to get conditions that capture target DNA with high sensitivity and selectivity. Parameters determining the performance of the microarray are spot morphology, ...

Microarray technology, like many other surface-capture diagnostic methods, relies on fidelity of affinity interactions between a surface-bound probe (e.g., nucleic acid or antibody) and its target in the sample milieu to produce an assay signal specific to analyte. These interfacial ...

A number of hetero- and homobifunctional reagents have been reported to immobilize biomolecules on a variety of supports. However, efforts are on to search for a method, which is relatively simple, involving minimum of steps, cost effective, easy to reproduce, and that produces stable oligon ...

Oligonucleotide microarray has become an important and powerful tool for various genomic analyses, where, unlike conventional methods, one can identify simultaneously a large number of targets in a given sample. Postsynthesis immobilization, the most widely used method, involv ...

A method of high resolution, array-based comparative genomic hybridization is described for the mapping of copy-number changes associated with chromosomal amplifications, deletions, and translocations. The method involves the design of whole-genome or targeted, fine-til ...

The mouse has become an important model for understanding human development, physiology and disease because of its genetic and biological similarity to humans. Desired mouse mutants with precise genetic alterations can now be generated through gene targeting in mouse embryonic stem ...

The use of genetically engineered mice has become a standard approach in order to study the physiological contribution of genes in a variety of life-science disciplines. Classical and conditional gene-targeting methods aimed at generating knock-out mice that lack gene products have b ...

Gene targeting in ES cells is extensively used to generate designed mouse mutants and to study gene function in vivo. Knockout mice that harbor a null allele in their germline provide appropriate genetic models of inherited diseases and often exhibit embryonic or early postnatal lethality. ...

Gene targeting in mouse embryonic stem cells is an essential, yet still very expensive and highly time-consuming, tool and method to study gene function at the organismal level or to create mouse models of human diseases. Conventional cloning-based methods have been largely used for genera ...

A culmination of large-scale ideas and efforts has truly allowed for the use of large genomic DNA clones housed in Bacterial Artificial Chromosome (BAC) vectors for biological research. Fundamental advances that have allowed this to happen include (1) the completion of genome sequencing ...

Recombinase-mediated cassette exchange (RMCE) is a powerful tool to generate a series of knock-in mutations into a particular gene of the mouse. It uses standard ES cell technologies to introduce an exchangeable cassette into the gene of interest by homologous recombination. Because the ...

Molecular genetic strategies to study gene function in mice or to generate a mouse model for a human disease are continuously under development. The application and importance of knock-in approaches are increasing. This chapter elaborates on novel developments for the generation of kn ...

Iodine-125 radioprobing is based on incorporation of radioiodine into a defined position in a nucleic acid molecule. Decay of 125I results in the emission of multiple, low-energy Auger electrons that, along with positively charged residual daughter nuclide, produce DNA strand breaks. ...

Fluorescent reporter groups have served for many years as sensitive probes of macromolecular structure. Such probes can be especially useful in comparative studies such as detection of conformational changes and discrimination among structural models. Spectroscopic metho ...

Analytical ultracentrifugation (AUC) is a powerful technique for the characterization of hydrodynamic and thermodynamic properties. The intent of this article is to demonstrate the utility of sedimentation velocity (SV) studies to obtain hydrodynamic information for G-qua ...