遗传学实验技术

Advancements in transgenic technologies have made the mouse one of the most useful animal models for biomedical research. Several technological breakthroughs have allowed the generation transgenic and knockout mouse models some 25 years ago. Subsequently, the technology has un ...

The generation of genetically modified animals using N-ethyl-N-nitrosourea (ENU) mutagenesis is a fast and highly effective method. The technique is based on treating male animals with the supermutagen ENU, which randomly introduces mutations in the spermatogonial stem cells. By b ...

A method for random relatively unbiased mutagenesis of ES cells with a mutagenic retroviral vector is described. An orderly assembly of mutant ES cells in multi-well plates is generated. 3D pooling of the wells of the assembly allows quick PCR search for insertions in genes of interest. Mutant ES ce ...

Traditionally, transgenic mice are generated by pronuclear injection of exogenous DNA. This technique has several limitations, including limited control over transgene expression, transgene cytotoxicity, �promiscuity and silencing, and founder mouse sterility. Here we ...

Recent advances in genome sequencing technology and algorithms have made it possible to determine the sequence of a whole genome quickly in a cost-effective manner. As a result, there are more than 200 completely sequenced genomes. However, annotation of a genome is still a challenging task. One ...

We describe BugView, a cross-platform application for presenting and comparing the genomes of bacteria or eukaryotes. We give particular emphasis to its use in comparing the genes of related bacterial genomes, and consider different methods of automating the preparation of genome com ...

Recent years have brought a tremendous increase in the amount of sequence data from various bacterial genome sequencing projects, an increase that is projected to accelerate over the next years. Comparative genomics of microbial strains has provided us with unprecedented informati ...

GenColors (gencolors.fli-leibniz.de) is a new web-based software/database system aimed at an improved and accelerated annotation of prokaryotic genomes considering information on related genomes and making extensive use of genome comparison. It offers a seamless integrat ...

Comparison of related genomes is an enormously powerful technique for explaining phenotypic differences and revealing recent evolutionary events. Genomes evolve through a host of mechanisms including long- and short-range intragenomic rearrangements, insertion of late ...

Guanine-rich sequences have been shown to readily form parallel or antiparallel G-quadruplex DNA structures in vitro. All telomeric repeat sequences contain stretches of guanine residues that can form quadruplex structures. In order to demonstrate the occurrence of the quadrupl ...

DNA oligonucleotides containing guanine repeat sequences can adopt G-quadruplex (GQ) structures in the presence of specific metal ions. We report on how to use a combination of spectroscopic and calorimetric techniques to determine the spectral characteristics and thermodyna ...

Comparative genome analysis is critical for the effective exploration of a rapidly growing number of complete and draft sequences for microbial genomes. The Integrated Microbial Genomes (IMG) system (img.jgi.doe.gov) has been developed as a community resource that provides suppo ...

Comparative analysis of DNA sequence from multiple species can provide insights into the function and evolutionary processes that shape genomes. The University of California Santa Cruz (UCSC) Genome Bioinformatics group has developed several tools and methodologies in its study ...

This chapter discusses VISTA Browser and associated computational tools for analysis and visual exploration of genomic alignments. The availability of massive amounts of genomic data produced by sequencing centers stimulated active development of computational tools for a ...

The availability of an increasing number of whole genome sequences presents us with the need for tools to quickly put them into a nucleotide-level multiple alignment. Mercator and MAVID are two programs that can be combined to accomplish this task. Given multiple whole genomes as input, Mercat ...

The Lagan Toolkit is a software package for comparison of genomic sequences. It includes the CHAOS local alignment program, LAGAN global alignment program for two, or more sequences and Shuffle-LAGAN, a “glocal” alignment method that handles genomic rearrangements in a global alignment ...

DIALIGN is a software program for multiple alignment of DNA or protein sequences that combines global and local alignment features. During the last years, the program has been used extensively to compare syntenic regions in genomic sequences. An anchoring option speeds up the alignment pr ...

In the current genomics era, protein and DNA sequence databases are continuously growing at an exponential rate. It has become increasingly important and useful to repeat similarity searches at frequent intervals, which then retrieve larger and larger sets of results. In addition, sequ ...

PSI-BLAST (Position-Specific Iterative Basic Local Alignment Search Tool) derives a position-specific scoring matrix (PSSM) or profile from the multiple sequence alignment of sequences detected above a given score threshold using protein–protein BLAST. This PSSM is used to fur ...

During the past few years, retroviral vectors have become a very important and widely used means of gene transfer. In the laboratory, their use has expanded the capabilities of investigators to perform important experiments that have solved previously unanswerable biological quest ...