生物化学技术

Recently, much attention has been focused on triplet-repeat expansions on the human genome, because they are reported to cause a number of neurodegenerative diseases such as the familial mental retardation, myotonic dystrophy, autosomal dominant diseases, or Huntington disease, ...

The high sensitivity of the reverse transcriptase-polymerase chain reaction (RT-PCR) allows the amplification of low abundance mRNA transcripts, but the biomedical research and diagnostics community have increasing interest in the ability to accurately quantify the amounts ...

The chemical mismatch cleavage (CMC) analysis was first described by Cotton et al. (1) and has successfully been used for detection and identification of mutations in several genes implicated in causing human genetic disorders (2-7). Compared with some current methods, such as denaturing ...

Differential display polymerase chain reaction (DD-PCR), originally described by Liang and Pardee in 1992 (1), has been utilized to study gene expression events in a myriad of biological processes. These processes include cell differentiation, hormonal regulation of gene express ...

It is possible to analyze RNA extracted from paraffin embedded-tissues that have been fixed in formalin (1-3). In fact, all biopsy or surgical tissues are routinely fixed and paraffin-embedded and conserved for many years for future examination, and present a very valuable medical and resea ...

The human genome initiative (1-3) has spurred the development of analytical instrumentation for high-throughput DNA sequencing. Originally carried out only in poly(acrylamide) slab gels, DNA sequencing is currently to an increasing extent performed in the capillary electroph ...

The Human Genome Project is a challenging and important project for the future of mankind (1). It has driven the development of various analytical instruments as well as methods (2). The development of an automated DNA sequencer with a fluorescence detection system has been a key factor for the succ ...

DNA analysis methods have been greatly developed in the past 10 yr with the progress of the Human Genome Project (1). In addition to rapid and high-throughput DNA sequencers (2), various reagents (3), and sample preparation methods (4-5)have been developed as well. In order to suit for large scale DNA se ...

Capillary electrophoresis (CE) has become an alternative to slab-gel electrophoresis for DNA separations due to its many advantages such as speed, increased separation efficiency, requires minute amount of sample, and automation of sample loading (1) Currently, high-throughput ...

Both directed and nondirected techniques are used in population biology for the identification of the genetic source of variation and the genetic locus of disease or quality traits (1,2). The “nondirected” approaches use random genome scanning methods initially to generate polymorp ...

Many human genetic diseases are caused by small alterations in DNA sequence of specific gene(s) (1,2. Many different types of DNA mutations and polymorphisms are found to contribute to the alterations in the DNA sequence of disease-causing genes. These polymorphisms include sequence cha ...

Described in this chapter is a capillary electrophoresis (CE) method for the accurate determination of protein-DNA binding constants (1). The advantages of the method include the fast separation and quantitation of protein-DNA complex and free DNA, in uncoated capillaries without si ...

The interaction of DNA with proteins is a central theme of molecular biology. Although all cells in the organism contain the same genetic “blueprint,” this information is utilized in a highly selective and specific manner. Different types of cells activate different subsets of genes. These p ...

We have been developing a method for the study of small-molecule interactions with DNA, typically with ligands of less than 1000 Dalton. Such interactions are of interest in biochemistry, where cell signaling may involve DNA at various stages, but the main driving force in our studies has been the p ...

Flow cytometry is literally measuring cells while moving in a liquid. More specifically, a suspension of single cells is labeled with one or several fluorescent labels. In the machine, the cells are constrained into single file. These cells pass through one or more laser beams to excite the fluore ...

Now that the human genome and the genomes of a growing number of important model eukaryotic organisms such as yeast and mouse have been sequenced, research emphasis in the “postgenomic” era is beginning to shift to events downstream of the whole genome. Interest is now focused on the identificati ...

Recent data released by the Human Genome Consortium and Celera Genomics estimates that the human genome might contain about 30,000 protein coding genes (1, and references therein), of which about 15‰ are believed to be expressed in any given cell type (see Chapter 42). The set of expressed proteins a ...

Microsatellites, also known as STRs (short tandem repeats), are tandem repeats of a simple dinucleotide, trinucleotide, tetranucleotide, pentanucleotide, or hexanucleotide sequence (two to six bases) that occur abundantly and at random throughout most eukaryotic genomes (ro ...

For historical background regarding karyotyping, chromosome nomenclature, and further details for some of the methodologies presented in this chapter, the reader might wish to consult a number of publications (1–5). Karyotyping is a process by which individual chromosomes are iden ...

Antibodies play a vital role in immune defense against pathogens. They are globular glycoproteins produced by plasma cells in response to the antigenic stimulation of B-lymphocytes. Their circulation in blood and lymph contributes to the humoral component of the vertebrate immune sy ...