One major goal of genetic research is to understand the role of genetic variation. By far the most common type of such variation in humans involves single DNA bases, and is termed single nucleotide polymorphism (SNP). In order that DNA chip technology continue to emerge as an alterative to conventional DNA diagnostic methods, questions about the conformation and activity of DNA on surfaces must be addressed. We developed three robust chemical methods for the covalent attachment of amino-modified oligonucleotides on glass surfaces based on polymeric coatings, providing robust platforms to prepare oligonucleotide arrays. We explored the feasibility of using oligonucleotide-array methods for molecular typing of a highly polymorphic genomic region by using the PCR-LDR strategy. We set up a low resolution microarray-based molecular tool for allele discrimination, which is of interest in clinical applications. Our goal was the identification of Single Nucleotide Polymorphisms in the Human LeukocyteAntigen gene. The methodology we proposed presents interesting features as genotyping tool: the reaction principle is robust, the assay easily automated, thereaction format flexible and capable to analyse different samples in parallel.
Development of DNA microarrays for human single nucleotide polymorphisms detection
CONSOLANDI, CLARISSA
2004
Abstract
One major goal of genetic research is to understand the role of genetic variation. By far the most common type of such variation in humans involves single DNA bases, and is termed single nucleotide polymorphism (SNP). In order that DNA chip technology continue to emerge as an alterative to conventional DNA diagnostic methods, questions about the conformation and activity of DNA on surfaces must be addressed. We developed three robust chemical methods for the covalent attachment of amino-modified oligonucleotides on glass surfaces based on polymeric coatings, providing robust platforms to prepare oligonucleotide arrays. We explored the feasibility of using oligonucleotide-array methods for molecular typing of a highly polymorphic genomic region by using the PCR-LDR strategy. We set up a low resolution microarray-based molecular tool for allele discrimination, which is of interest in clinical applications. Our goal was the identification of Single Nucleotide Polymorphisms in the Human LeukocyteAntigen gene. The methodology we proposed presents interesting features as genotyping tool: the reaction principle is robust, the assay easily automated, thereaction format flexible and capable to analyse different samples in parallel.I documenti in UNITESI sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
https://hdl.handle.net/20.500.14242/78048
URN:NBN:IT:UNIMI-78048