遗传学实验技术

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 ...

Agrobacterium-mediated transformation of wheat is becoming a viable alternative to the more established biolistic protocols. It offers advantages in terms of simple, low-copy-number integrations and can be applied with similar efficiencies to specific durum wheat and spring a ...

The ability to use hCFs as vectors for introducing large stretches of human DNA into mice was first demonstrated in 1997 (1). Transferred hCFs were stably maintained as an extra chromosome in the somatic cells of mice, and the human genes included in them were expressed under proper tissue-specific ...

Amethod to achieve the insertion of genetic material into the chromosome of Escherichia coli is described. The method is based on the use of integration vectors from the pBRINTs-rAnbR family. These vectors offer the choice of using the antibiotics chloramphenicol, gentamycin, or kanamy ...

Successful transplantation of tissue during solid organ and bone marrow transplantation relies on accurate determination of the human leukocyte antigen (HLA) phenotype of the potential donor(s) and recipient. Matching donor with recipient for a kidney transplant generally mea ...

High combinatorial phage display libraries have become an important tool in the search for ligand-receptor interactions. The advantage this approach offers is the ability to screen large repertoires of peptides, displayed on the coat proteins of bacteriophages, against a target at the ...

Sensitive methods for the quantification of DNA fragments can be used to study an individual’s genetic constitution for duplicated regions of the genome or to determine the relative proportion of different DNA fragments in heterogeneous samples such as pooled DNA from different indiv ...

Single-nucleotide polymorphisms (SNPs) are common in the human genome, with more than 11 million SNPs having frequencies greater than 1%. The challenge is to identify the minority of functional SNPs from the large number of SNPs that are expected to be silent. Whereas coding variants are unusua ...

Several statistical tests have been introduced for the comparison of serial analysis of gene expression (SAGE) libraries to quantitatively analyze the differential expression of genes. As each SAGE library is only one measurement, the necessary information on biological variat ...