遗传学实验技术

The sperm cell has evolved to transmit a paternal haploid genome to the oocyte and form a new embryo. Therefore, it is essential that the integrity of this genome be evaluated as part of the standard semen analysis. The assessment of DNA fragmentation is consequently considered as an important para ...

The ability to non-invasively assess DNA oxidation and its repair, has significant utility in large-scale, population-based studies. Such studies could include the assessments of: the efficacy of antioxidant intervention strategies, pathological roles of DNA oxidation in vari ...

The binding of chemical carcinogens to DNA is well established as the initiating step in the process of carcinogenesis. While early studies in animals or cells in culture took advantage of radiolabeled model carcinogens such as benzo(a)pyrene, interest in measuring DNA damage levels in hum ...

Measurement of DNA double-strand break (DSB) levels in cells is useful in many research areas, including those related to DNA damage and repair, tumorigenesis, anti-cancer drug development, apoptosis, radiobiology, environmental effects, and aging, as well as in the clinic. DSBs can be det ...

The Buccal Micronucleus Cytome (BMCyt) assay is a new minimally invasive system for studying DNA damage, chromosomal instability, cell death, and the regenerative potential of buccal mucosal tissue. This method is increasingly being used in molecular epidemiologic studies inves ...

The cytokinesis-block micronucleus cytome (CBMN cyt) assay is a new and comprehensive technique for measuring DNA damage, cytostasis, and cytotoxicity in different tissue types, including lymphocytes. DNA damage events are scored specifically in once-divided binucleated ce ...

Enzyme-Linked Immuno Spot (ELISpot) assay is widely used for vaccine development, cancer and AIDS research, and autoimmune disease studies. The output of an ELISpot assay is a formation of colored spots which appear at the sites of cells releasing cytokines, with each individual spot repres ...

Exocyclic etheno–DNA adducts are formed by the reaction of lipid peroxidation products, such as 4-hydroxy-2-nonenal (HNE) with DNA bases to yield 1,N 6-etheno-2′-deoxyadenosine (εdA), 3,�N 4-etheno-2′-deoxycytidine (εdC), and etheno-2′-deoxyguanosine. These adducts act as a drivi ...

Co-immunoprecipitation (Co-IP) (followed by immunoblotting) is a technique widely used to characterize specific protein–protein interactions. Investigating interactions of proteins containing “sticky” polyalanine (PolyA) tracts encounters difficulties using ...

PCR amplification (followed by mutation scanning or direct sequencing) is a technique widely used in mutation detection and molecular studies of disease-causing genes, such as ARX. PCR amplification of high GC-rich regions encounters difficulties using conventional PCR proced ...

Fragile X syndrome, the leading inherited cause of mental retardation and autism spectrum disorders worldwide, is caused by a tandem repeat expansion in the FMR1 (fragile X mental retardation 1) gene. It presents with a distinct behavioral phenotype which overlaps significantly with th ...

Pulse shape analysis (PulSA) is a flow cytometry-based method that can be used to study protein localization patterns in cells. Examples for its use include tracking the formation of inclusion bodies of polyglutamine-expanded proteins and other aggregating proteins. The method can al ...

The polyglutamine diseases are caused by the expansion of CAG repeats. A key step in understanding the disease mechanisms, at the DNA and protein level, is the ability to produce recombinant proteins with specific length glutamine tracts which is a time-consuming first step in setting up in vitro ...

Defining the aggregation process of proteins formed by poly-amino acid repeats in cells remains a challenging task due to a lack of robust techniques for their isolation and quantitation. Sedimentation velocity methodology using fluorescence detected analytical ultracentri ...

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder. The HD gene encodes the huntingtin protein (HTT) that contains polyglutamine tracts of variable length. Expansions of the CAG repeat near the amino terminus to encode 40 or more glutamines (polyQ) lead to dis ...

Mutations which cause an expansion of CAG triplet repeats encoding polyglutamine (polyQ) are responsible for the subsequent misfolding of specific proteins that contribute directly to the pathogenesis of at least nine neurodegenerative disorders, including Huntington’s d ...

Misfolded proteins have been implicated in most of the major neurodegenerative diseases, and identifying drugs and pathways that protect neurons from the toxicity of misfolded proteins is of paramount importance. We invented a form of automated imaging and analysis called robotic mi ...

Spinal and bulbar muscular atrophy (SBMA) is a late-onset neurodegenerative disease caused by a polyglutamine expansion in the androgen receptor (AR). In vivo and in vitro studies have suggested that some steps of normal AR function and metabolism, such as hormone binding and nuclear trans ...

Aberrant expansion of the number of polyglutamine (polyQ) repeats in mutant proteins is the hallmark of various diseases. These pathologies include Huntington’s disease (HD), a neurological disorder caused by expanded polyQ stretch within the huntingtin (Htt) protein. The expans ...

Numerous proteins contain domains that are enriched in glutamine and asparagine residues, and aggregation of some of these proteins has been linked to both prion formation in yeast and a number of human diseases. Unfortunately, predicting whether a given glutamine/asparagine-rich p ...