Rapid, high fidelity analysis of simple sequence repeats on an electronically active DNA microchip
AUTOR(ES)
Radtkey, Ray
FONTE
Oxford University Press
RESUMO
We describe a method for the discrimination of short tandem repeat (STR) alleles based on active microarray hybridization. An essential factor in this method is electronic hybridization of the target DNA, at high stringency, in <5 min. High stringency is critical to avoid slippage of hybrids along repeat tracts at allele-specific test sites in the array. These conditions are attainable only with hybridization kinetics realized by electronic concentration of DNA. A sandwich hybrid is assembled, in which proper base stacking of juxtaposed terminal nucleotides results in a thermodynamically favored complex. The increased stability of this complex relative to non-stacked termini and/or base pair mismatches is used to determine the identification of STR alleles. This method is capable of simultaneous and precise identification of alleles containing different numbers of repeats, as well as mutations within these repeats. Given the throughput capabilities of microarrays our system has the potential to enhance the use of microsatellites in forensic criminology, diagnostics and genetic mapping.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=102800Documentos Relacionados
- A simple, rapid, high-fidelity and cost-effective PCR-based two-step DNA synthesis method for long gene sequences
- Rapid, Simple In Vivo Screen for New Drugs Active against Mycobacterium tuberculosis
- Rapid, high-level expression of biologically active alpha-trichosanthin in transfected plants by an RNA viral vector.
- Rapid, Simple Enzyme Immunoassay for Gentamicin
- A rapid, simple assay for digoxin