遗传学实验技术

The central cause to any cancer ultimately lies in the genome and the initial alterations that result in changes in gene expression that are reflected in the phenotype of the cancer cell. The gene expression data are rich in information but the primary lesions responsible for carcinogenesis are ...

Many oligonucleotide arrays comprise of spotted short oligonucleotides from throughout the genome under study. Hybridization of tumor DNA samples to these arrays will provide copy number estimates at each reference point with varying degrees of accuracy. In addition to copy number c ...

Molecular inversion probe (MIP) technology has been demonstrated to be a robust platform for large-scale dual genotyping and copy number analysis. Applications in human genomic and genetic studies include the possibility of running dual germline genotyping and combined copy numb ...

High-resolution genomic arrays and next-generation sequencers are some of the genome-based technologies poised to make significant contributions in the near future to basic and clinical science. The success of these technologies, and most certainly their translation into the cl ...

Differential methylation hybridization (DMH) is a high-throughput DNA methylation screening tool that utilizes methylation-sensitive restriction enzymes to profile methylated fragments by hybridizing them to a CpG island microarray. This array contains probes spanni ...

In this chapter, we introduce a few statistical algorithms for calling gains and losses in array-based comparative genomic hybridization (array CGH) data, including CBS, CLAC, CGHseg, and Fused Lasso. We illustrate the performance of the methods through simulated and real data examples. ...

Transgenic animal models are valuable tools for testing gene functions and drug mechanisms in vivo. They are also the best similitude of a human body for etiological and pathological research of diseases. All pharmaceutically developed drugs must be proven safe and effective in animals be ...

When trying to obtain a cDNA clone to a novel protein, the only handle one may have is an antibody that recognizes the protein of interest. Consequently, the obvious approach is to screen a λ-phage expression library using the antibody. In general, polyclonal sera give better results, but a mixture of mo ...

T-cell clonal expansion has been detected in the peripheral blood or the disease-affected sites in patients who suffer from various disease states such as infections, autoimmune diseases, malignancy, and post-transplantation complications (1–7). Since T-cells begin to prolifer ...

Faithful gene activity reporters are a useful tool for evo-devo studies enabling selective introduction of specific loci between species and assaying the activity of large gene regulatory sequences. The use of large genomic constructs such as BACs and fosmids provides an efficient pla ...

Genomic DNA libraries are a valuable source of large constructs that can contain all the regulatory elements necessary for recapitulating wild-type gene expression when introduced into animal genomes as a transgene. Such clones can be directly used in complementation studies. In com ...

Nucleic acid arrays provide a powerful methodology for studying biological systems on a genomic scale. BD Atlas™ Arrays, developed by BD Biosciences Clontech, are expression profiling products specifically designed to be accessible to all laboratories performing isotopic blot h ...

Since DNA topoisomerases have become a major focus for scientists, the purification of these proteins from all sources represents one of the basic hurdles on the way to investigating this important class of enzymes. A variety of methods consisting of numerous steps have been applied to obtain a r ...

Current cancer chemotherapy strategies are often hampered by the lack of tumor selectivity, resulting in unwanted damage to healthy tissue. Gene-directed enzyme-prodrug therapy (GDEPT) (1) and virus-directed enzyme-prodrug therapy (VDEPT) (2) are suicide gene therapy approac ...

The postreplicative methylation of DNA at the C5 position of cytosines is found in a broad spectrum of organisms ranging from prokaryotes to human (1). In prokaryotes the major role of cytosine C5 methylation (like adenine N6 and cytosine N4 methylation) is to protect the genome against DNA degra ...

Apoptosis, frequently referred to as “programmed cell death” or “cell suicide,” is an active and physiological mode of cell death, where the cell itself prepares and executes the program of its own demise. A complex, multistep mechanism regulates the cell’s response to various stimuli by apopt ...

The full-length cDNA, which contains the entire sequence of the mRNA, is the ultimate goal for cDNA cloning. Unfortunately, cDNA libraries constructed by many types of conventional methods have a high content of nonfull-length cDNA clones. One of the reasons for this is that reverse transcrip ...

Systematic analysis of an organism’s transcript repertoire plays a key role in molecular genetics. The complexity of this task in higher organisms such as mammals is not only owing to the large number of expected genes (70,000–100,000) in humans (1) but also because gene coding regions are disper ...

One of the greatest improvements in the use of clone libraries in genomic research was the introduction of library arrays by storing single clones in separate wells of microtiter plates. This not only makes clones practically immortal by keeping the plates at −80�C, but it also gives each of the clon ...

During the last decade, major advances in genomic research have resulted in the development of a variety of tools and technologies for gene identification that were successfully applied to the construction of transcriptional maps and to the identification of genes responsible for gen ...