PMID: 7526344Oct 25, 1994Paper

High-density multiplex detection of nucleic acid sequences: oligonucleotide ligation assay and sequence-coded separation

Nucleic Acids Research
P GrossmanD A Iovannisci

Abstract

We describe a non-isotopic, semi-automated method for large-scale multiplex analysis of nucleic acid sequences, using the cystic fibrosis transmembrane regulator (CFTR) gene as an example. Products of a multiplex oligonucleotide ligation assay (OLA) are resolved electrophoretically from one another and from unligated probes under denaturing conditions with fluorescence detection. One ligation probe for each OLA target carries a fluorescent tag, while the other probe carries an oligomeric non-nucleotide mobility modifier. Each OLA product has a unique electrophoretic mobility determined by the ligated oligonucleotides and the mobility-modifier oligomer arbitrarily assigned (coded) to its target. The mobility range for practical mobility modifiers is much wider than the accessible range from unmodified ligated oligonucleotides of practical length. Each mobility modifier is built from phosphoramidite monomers in a stepwise manner on its associated oligonucleotide using an automated synthesizer. The resulting mobility modifiers lower the probe-target duplex Tm by less than 3 degrees C and retard probe-target annealing by less than 50%, with negligible effect on OLA yield and specificity. This method is especially useful for allelic...Continue Reading

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Citations

Jan 5, 2002·Early Human Development·M A Gómez-LlorenteJ A Gómez-Capilla
Feb 13, 2001·Current Opinion in Biotechnology·J BanérU Landegren
Dec 29, 2005·Genetic Testing·Pietro StanzialeLuigi Bisceglia
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Jun 18, 2009·Nucleic Acids Research·Marco SevergniniBianca Castiglioni
Aug 14, 2010·Genetics in Medicine : Official Journal of the American College of Medical Genetics·Marco LucarelliSerena Quattrucci
Jul 20, 2010·Annual Review of Analytical Chemistry·Tim ConzeUlf Landegren
Sep 21, 2005·BMC Bioinformatics·Johan StenbergUlf Landegren
Dec 17, 2005·Genome Biology·Stuart J MacdonaldAnthony D Long
Sep 25, 1999·Journal of Neuroimmunology·J ChatawayA Compston
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Sep 27, 2005·Forensic Science International : Synergy·Beatriz SobrinoAngel Carracedo
Apr 26, 2006·Analytical Biochemistry·Marco LucarelliRoberto Strom
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Nov 4, 2017·PloS One·Martín E MautnerDaniel Corach
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Aug 30, 2003·Archives of Pathology & Laboratory Medicine·Elaine Lyon, Christine Miller
Sep 14, 2018·Biomacromolecules·Xun HanChristopher N Bowman

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