			Home About us Contact

Mutation Scanning (mutation + scanning)

Distribution by Scientific Domains

Life Sciences	87%

Selected Abstracts

Description and validation of high-throughput simultaneous genotyping and mutation scanning by high-resolution melting curve analysis,

HUMAN MUTATION, Issue 6 2009
Tú Nguyen-Dumont
Abstract Mutation scanning using high-resolution melting curve analysis (HR-melt) is an effective and sensitive method to detect sequence variations. However, the presence of a common SNP within a mutation scanning amplicon may considerably complicate the interpretation of results and increase the number of samples flagged for sequencing by interfering with the clustering of samples according to melting profiles. A protocol describing simultaneous high-resolution gene scanning and genotyping has been reported. Here, we show that it can improve the sensitivity and the efficiency of large-scale case,control mutation screening. Two exons of ATM, both containing an SNP interfering with standard mutation scanning, were selected for screening of 1,356 subjects from an international breast cancer genetics study. Asymmetric PCR was performed in the presence of an SNP-specific unlabeled probe. Stratification of the samples according to their probe-target melting was aided by customized HR-melt software. This approach improved identification of rare known and unknown variants, while dramatically reducing the sequencing effort. It even allowed genotyping of tandem SNPs using a single probe. Hence, HR-melt is a rapid, efficient, and cost-effective tool that can be used for high-throughput mutation screening for research, as well as for molecular diagnostic and clinical purposes.Hum Mutat 30:1,7, 2009. © 2009 Wiley-Liss, Inc. [source]

Cover Picture: Electrophoresis 16'09

ELECTROPHORESIS, Issue 16 2009
Article first published online: 18 AUG 200
Issue no. 16 is a special on "Enantioseparations". It consists of 19 research papers and 2 review articles distributed over 4 different parts. The two review articles make up Part I and focus on recent developments in microchip enantioseparations and chiral analysis of drugs, metabolites and biomarkers in biological samples. The 19 research papers are distributed over the remaining 3 parts including "Fundamentals and Methodologies", "Chiral Capillary Electrochromatography" and "Biomedical, Pharmaceutical, Food and Environmental Applications of Electromigration Techniques". Issue no. 16 also includes a Fast Track paper on the "Analysis of genetic variation in Globocephaloides populations from macropodid marsupials using a mutation scanning-based approach". [source]

Quantification of sequence exchange events between PMS2 and PMS2CL provides a basis for improved mutation scanning of lynch syndrome patients,

HUMAN MUTATION, Issue 5 2010
Heleen M. van der Klift
Abstract Heterozygous mutations in PMS2 are involved in Lynch syndrome, whereas biallelic mutations are found in Constitutional mismatch repair-deficiency syndrome patients. Mutation detection is complicated by the occurrence of sequence exchange events between the duplicated regions of PMS2 and PMS2CL. We investigated the frequency of such events with a nonspecific polymerase chain reaction (PCR) strategy, coamplifying both PMS2 and PMS2CL sequences. This allowed us to score ratios between gene and pseudogene-specific nucleotides at 29 PSV sites from exon 11 to the end of the gene. We found sequence transfer at all investigated PSVs from intron 12 to the 3, end of the gene in 4 to 52% of DNA samples. Overall, sequence exchange between PMS2 and PMS2CL was observed in 69% (83/120) of individuals. We demonstrate that mutation scanning with PMS2 -specific PCR primers and MLPA probes, designed on PSVs, in the 3, duplicated region is unreliable, and present an RNA-based mutation detection strategy to improve reliability. Using this strategy, we found 19 different putative pathogenic PMS2 mutations. Four of these (21%) are lying in the region with frequent sequence transfer and are missed or called incorrectly as homozygous with several PSV-based mutation detection methods. Hum Mutat 31:578,587, 2010. © 2010 Wiley-Liss, Inc. [source]

Description and validation of high-throughput simultaneous genotyping and mutation scanning by high-resolution melting curve analysis,

Determination of the mutation spectrum of the EXT1/EXT2 genes in British Caucasian patients with multiple osteochondromas, and exclusion of six candidate genes in EXT negative cases,,

HUMAN MUTATION, Issue 11 2006
Lorne Lonie
Abstract We describe here the spectrum and distribution of mutations in the EXT1 and EXT2 genes in the largest reported British Caucasian multiple osteochondromas (MO) population. Furthermore, we report for the first time the screening of the EXT1 and EXT2 promoters, 5,UTRs, and 3,UTRs, and exclude six potential MO candidate genes in individuals without a detectable mutation within the coding region of EXT1 and EXT2. The coding exons of EXT1 and EXT2 were screened in 72 unrelated probands affected with MO. Forty-six different mutations were identified in 56 probands, of which 29 were novel. Mutation in the EXT1 and EXT2 genes each accounted for 50% of the mutations identified. Of the 72 probands, 42 were of British Caucasian descent, which when added to the 41 British Caucasian families previously reported from our total cohort, gave a total of 83 families. This cohort's proportional frequency for EXT1/EXT2 mutation was 53%/47%. We also validated the technique of high-resolution melting analysis in a blind study using 27 unique EXT1 or EXT2 mutations. This technique was found to be sensitive with a detection rate of 100% regarding heterozygote detection for EXT mutation scanning. Furthermore, this technique has a very high throughput and is very cost-effective. © 2006 Wiley-Liss, Inc. [source]

Rapid denaturing high-performance liquid chromatography (DHPLC) for mutation scanning of the transforming growth factor ,3 gene using a novel proof-reading polymerase

INTERNATIONAL JOURNAL OF IMMUNOGENETICS, Issue 5 2003
A. Bayat
Summary We have utilized a novel variation on the conventional denaturing high-performance liquid chromatography (DHPLC) technology, which we term rapid DHPLC, combining changes in instrumentation, cartridge technology and analysis conditions to enable significant increases in throughput to be achieved. In addition, the use of a novel proof-reading polymerase for sample amplification with a low misincorporation rate enables simplification of the DHPLC patterns and hence enhanced mutation detection recognition. This scheme for increasing DHPLC throughput has been tested by scanning the transforming growth factor (TGF) ,3 gene for the presence of mutations for which there is limited published or on-line data available regarding the presence of gene polymorphisms. TGF, isoforms have multiple roles in cell division, growth, proliferation, transformation and differentiation. TGF,3 is a TGF, cytokine isoform, and has an important role in embryogenesis, cell differentiation and wound healing. The TGF,3 gene consists of seven exons and six introns spanning 43 000 bp of the human genome on chromosome 14q23,24. The rapid DHPLC approach enabled scanning of all seven exons and part of the promoter region (1000 bp upstream from exon 1 in the 5,-flanking regions) of the TGF,3 gene in 95 Caucasian individuals in only 8 days, in comparison to the 17 days it would have previously taken. Mutations were clearly identified in the promoter region of the TGF,3 gene but were absent from the exonic regions. Understanding the genetic variations affecting the TGF,3 gene is important as this molecule has multiple regulatory functions on a variety of cell types. [source]