DNA-based measurement of copy number can be useful in a number of applications in human genetics, through its detection of rearrangements of chromosomal structure too small to be detected by standard cytogenetic analyses. In some cases, deletions or duplications may involve several lin ...

Automated fluorescent sequencing of polymerase chain reaction (PCR) products is now widely used in molecular diagnostics. It is most commonly used to characterize mutations detected in an initial screen using a less sensitive, indirect method, rather than a mutation scanning techni ...

Automated fluorescent sequencing is often overlooked as a point mutation scanning technique usually on the grounds of cost, technical complexity, and difficulties in processing the information generated. However, sequencing cannot be avoided completely, because regardle ...

The protein truncation test (PTT) is a powerful mutation detection technique originally described for mutation screening of the dystrophin gene (1). It is based on in vitro transcription/translation technology, and is capable of directly scanning kilobase-sized fragments of DNA for ...

This chapter describes the Transgenomic WAVE� DNA Fragment Analysis System with DNA Sep� Technology, as used in our molecular genetics diagnostic laboratories for the detection of unknown mutations. Four software packages are currently available: WAVEmaker 3.4, 4, 4.1, and most recent ...

The polymerase chain reaction (PCR) has rapidly become an essential tool within the diagnostic laboratory. Therefore, it is crucial when setting up a new PCR-based test to ensure that the PCR reaction is carefully designed to be as robust and reliable as possible. Usually, little optimization is ...

Genetic testing is often regarded as a laboratory procedure in a molecular genetic laboratory. However, for practical use in health care, this definition is too narrow. In the majority of cases, the clinician is not using a genetic test for diagnostic purposes, but confirming or excluding a clini ...

In the last decade, the demand for molecular genetic testing has increased enormously. Many of the laboratories that offer diagnostic molecular genetic testing on a routine basis originated from a research-based setting. The exponential growth of the clinical diagnostic molecular g ...

The spinal muscular atrophies are a clinically and genetically heterogeneous group of neuromuscular disorders caused by degeneration of anterior horn cells. In proximal spinal muscular atrophy (SMA), the muscles of the extremities closest to the trunk are affected earlier and more s ...

The two forms of dystrophin-associated muscular dystrophies, known as Duchenne and Becker muscular dystrophy (DMD/BMD; OMIM 310200) are caused by genetic defects in the huge DMD gene (79 exons), located at Xp21 and coding for the 427-kDa protein known as dystrophin (1,2). DMD is the most frequent m ...

Neurofibromatosis type 1 (NF1) is one of the most common dominantly inherited neurogenetic disorders, affecting about 1 in every 4000 individuals worldwide. It is fully penetrant by the age of five. The condition is characterized by multiple caf�-au-lait spots, benign neurofibromas, and ...

Multiple endocrine neoplasia (MEN) is characterized by the occurrence of tumors involving two or more endocrine glands within a single patient. Multiple endocrine neoplasia types 1 (MEN1) and 2 (MEN2) are autosomal dominant cancer syndromes caused by mutations in the MEN1 (1) and RET (2) gene ...

Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene (1,2). More than 1000 disease mutations and 200 polymorphisms have been identified in the CFTR gene (3). The type and frequency of mutations is very variable from ethnic popula ...

The hemoglobinopathies are a diverse group of inherited recessive disorders that include the thalassemias and sickle-cell disease. These were the first genetic diseases to be characterized at the molecular level, and consequently have been used as a prototype for the development of new ...

Since the discovery of dynamic mutations approx 10 yr ago, the pathological expansion of unstable trinucleotide repeats has been shown to be the cause of an increasing number of neurological disorders. The trinucleotide repeat disorders identified to date can be categorized into two sub ...

Familial adenomatous polyposis (FAP) is an autosomal, dominantly inherited disorder that predisposes to the development of hundreds to thousands of adenomatous polyps throughout the colon and rectum (MIM 175100). Left untreated, it is almost 100% certain that at least one of these polyps ...

Mapping of protein binding sites within the genome has been significantly advanced by microarray and sequencing technologies, yet the method traditionally used to isolate protein–DNA complexes, chromatin immunoprecipitation, has remained dependent of the use of antibodies. ...

Comprehensive analysis of DNA–protein interactions is important for mapping transcriptional regulatory networks at the genome-wide level. Here, we present a new application of mRNA display, using the in vitro virus (IVV) technology, for in vitro selection of DNA-binding protein com ...

Promoter deletion analysis is a useful tool for identifying important regulatory regions involved in transcriptional control of gene expression. In this approach, a series of promoter deletion fragments are fused to a reporter gene, such as chloramphenicol acetyltransferase or l ...

Luciferase based assays have become an invaluable tool for the analysis of cloned promoter DNA fragments, both for verifying the ability of a potential promoter fragment to drive the expression of a luciferase reporter gene in various cellular contexts, and for dissecting binding elemen ...