The occurrence of autoimmunity is strongly associated with multiple gene variants that predispose individuals to disease. The identification of the gene polymorphisms that modulate disease susceptibility is key to our understanding of disease etiology and pathogenesis. Whi ...

RNA interference is a potent gene silencing pathway initiated by short molecules of double-stranded RNA. Small interfering RNAs (siRNAs) with full sequence complementarity to mRNAs induce cleavage of their target transcripts in the cytoplasm. Recent evidence has shown, however, th ...

With the successful completion of genome sequencing projects for a variety of model organisms, the selection of candidate organisms for future sequencing efforts has been guided increasingly by a desire to enable comparative genomics. This trend has both depended on and encouraged the d ...

There are innumerable instances when one wants to manipulate cloned DNA: point mutagenesis for analysis of protein function and transcription factor binding sites, introduction of insertions to produce fusion proteins or to introduce reporter genes or motifs, and creation of delet ...

Flow cytometry and fluorescence-activated cell sorting (FACS) are techniques of great power used to screen cells rapidly for expression of particular gene products. These techniques have been of general utility in identifying and selecting populations of cells of defined charact ...

In situ polymerase chain reaction (PCR) refers to the amplification of specific nucleic acid sequences and subsequent visualization of the PCR products in tissue sections. When PCR is performed in fixed cells in suspension the term in-cell PCR is normally applied instead. For example, in-ce ...

Chromosome “painting” by fluorescent labeled probes for each human chromosome has been available since the late 1980s. The DNA or RNA radioactive isotopes have been in use since 1969 as nucleic acid probes (1,2). The introduction of fluorescent labeled probes by Bauman et al. (1980) presented an ...

Lentiviruses, such as human immunodeficiency virus (HIV), feline immunodeficiency virus (FIV), equine infectious anemia virus (EIAV), and bovine Jembrana disease virus (JDV) are members of the Retroviridae, viruses with enveloped capsids and a plus-stranded RNA genome. Like all re ...

Since their original description, liposomes have been discussed as vehicles that could be used as carriers of pharmaceutically active agents (1), and their potential for use as carriers of genetic information has been examined (2,3). Some encouraging DNA-delivery results have been obt ...

In situ hybridization is the only molecular biology based test that allows for the direct correlation of the results with the histologic and cytologic features of the sample. The DNA/RNA extraction that precedes filter hybridization (slot blot or Southern blot) and PCR precludes this anal ...

Complex biological fluids, such as the vast and molecularly crowded cell cytoplasm and the highly viscoelastic mucus that protects many entry ways to the body, pose significant barriers to efficient gene delivery. Understanding the dynamics of gene carriers in such environments allo ...

Since the discovery of DNA fingerprinting ten years ago (1), the direct analysis of DNA variation has been successfully used in a wide range of applications and has become an important tool in forensic science (2,3). Most DNA typing systems assay DNA fragment length at repeated loci showing tandem r ...

The polymerase chain reaction (PCR) is a method of central importance in molecular biology (1,2). The DNA fragment of interest is often amplified for cloning purposes. A frequently used experimental approach is to include extra restriction endonuclease cleavage sites in the amplifica ...

Using Escherichia coli cDNA microarray slides and Affymetrix GeneChips, we study how the relative position of probes on microarrays affects the cross-correlation of gene expression profiles. We find that in cDNA arrays, every spot located within the same block is affected by a similar, exp ...

Since the first report on cDNA cloning in 1972 (1), this technology has been developed into a powerful and universal tool in isolation, characterization, and analysis of both eukaryotic and prokaryotic genes. But the conventional methods of cDNA cloning require much effort to generate a libr ...

Genomic imprinting, though most extensively studied in mammals, has long been known to perform an important role in seed development in flowering plants. In this chapter, an overview of what is known to date about genomic imprinting in flowering plants and how this knowledge came into being will be ...

Denaturing high performance liquid chromatography (dHPLC) is a fast and reliable technique for the DNA variation screening (1,2).It can detect in minutes with close to 100% sensitivity and specificity single-base substitutions as well as small deletions and insertions in DNA fragmen ...

Single-nucleotide polymorphism (SNP) is a new term for an old concept. Geneticists have been trying for decades to find the genetic differences among individuals. Originally phenotypes were used, then protein sequence, electrophoresis, restriction fragment polymorphisms (R ...

A greater understanding of the molecular, biochemical, and genetic factors involved in the progression of a specific disease state has led to the development of genetic therapies using direct gene transfer to ameliorate the disease condition or correct a genetic defect in situ. Effective ...

A method is described for the floral transformation of wheat using a protocol similar to the floral dip of Arabidopsis. This method does not employ tissue culture of dissected embryos, but instead pre-anthesis spikes with clipped florets at the early, mid to late uninucleate microspore stage a ...