Calcium phosphate mediated transfection of genomic DNA samples into mammalian recipient cells can be used to isolate specific genes of interest. In theory, for any phenotype for which there is a suitable selection in cell culture, a gene sequence encoding that phenotype can be transferred a ...

Evaluation of single-locus probe results in criminal-identity cases is best presented within the standard forensic framework for handling trace evidence. A comprehensive account was given by Aitken (1), and the language of that text will be used here. For simplicity, suppose a stain from the ...

Since its discovery by Prof. Sir Alec Jeffreys FRS in 1984, DNA fingerprinting using multilocus VNTR probes (MLPs) has been widely used in the field of human identification (1,2). Although multilocus probes were used in forensic investigations in the late 1980s (3), forensic samples often yield ...

PCR-VNTRs are important markers for questions of identification, individualization, and discrimination. They already form an integral part of forensic DNA analysis. They can be subdivided into the systems of short tandem repeats (STR) with fragment lengths ranging between approx 1 ...

Highly polymorphic variable number tandem repeat (VNTR) loci have proven very useful for human DNA testing purposes (1-3). Initially, these variants were characterized by restriction fragment-length polymorphism (RFLP) analysis. Subsequently, the polymerase chain reacti ...

PCR optimization is usually performed in order to obtain maximum specificity and yield. In some applications for which the amount of template may be limiting, or when there is a large amount of nontarget sequences, the sensitivity is maximized. Nonoptimized conditions promote artifactu ...

Short tandem repeat (STR) loci consist of repetitive elements of 3-7 nucleotides. The STR loci, which are numerous in the human genome, are highly polymorphic in length and may also vary in the sequences of the repetitive elements. The polymerase chain reaction (PCR) makes it possible to analyze ve ...

This introduction is intended to inform the forensic scientist of the historical development of STR typing on automated sequencers, as these precedents will have a bearing on the techniques and equipment used in the present-day laboratory. The section then concludes with a brief outline of ...

This chapter describes the manual typing of STR markers, detailing the electrophoretic techniques used to separate amplified products, and the relevant methods for their detection.

Mitochondrial DNA (mtDNA) typing is increasingly used for the forensic identification of human remains (1-9). This is especially true when only limited quantities of sample are present, such as when the sample has undergone extensive degradation and nuclear-typing methods are ineffe ...

For separation of alleles of PCR-dependent short tandem repeat (STR) polymorphisms, high resolution with the power to discriminate fragments differing by one base pair is often desired. Furthermore, sequencing reactions also require unambiguous one base-pair separation. For bo ...

The isolation of genomic deoxyribonucleic acid (DNA) is a crucial step in the process of DNA profiling. The success of all subsequent genetic-typing procedures depends on the availability of sufficient amounts of highly purified DNA from biological crime stains as well as from reference b ...

Oat (Avena sativa L.), a worldwide temperate cereal crop, is deficient in tolerance to osmotic stress due to drought and/or salinity. To genetically transform the available commercial oat cultivars, a genotype-independent and efficient regeneration system from shoot apical merist ...

Ever since the first developments in plant transformation technology using model plant species in the early 1980s, there has been a body of plant science research devoted to adapting these techniques to the transformation of crop plants. For some crop species progress was relatively rapid, ...

The low frequency and randomness of transgene integration into host cells, combined with the significant challenges of recovering whole plants from those rare events, makes the use of selectable marker genes routine in plant transformation experiments. For research applications ...

We present a complete, step-by-step guide to the production of transformed wheat plants using a particle bombardment device to deliver plasmid DNA into immature embryos and the regeneration of transgenic plants via somatic embryogenesis. Currently, this is the most commonly used meth ...

Reporter genes have been widely used in plant molecular biology, typically to discern patterns of gene expression, but also as markers of transformed cells during stable transformation procedures. The ideal marker gene would be expected to display characteristics such as ease and chea ...

The engineering of novel properties and functions into viral vectors for improved gene delivery remains a barrier to the development of efficient, customized gene delivery vehicles. Rational methods for designing improved viral vectors are often experimentally challenging and ...

Photochemical internalization (PCI) is a physico-chemical targeting method that enables light directed delivery of nucleic acids into cells. The technology is based on photosensitizers that localize in the membranes of endocytic vesicles. A light activation of the photosensit ...

The recent development of leukemia in gene therapy patients with X-linked severe combined immunodeficiency disease because of retroviral vector insertional mutagenesis has prompted reassessment of the genotoxic potential of integrating vector systems. In this chapter, va ...