title: G-quadruplex-generating polymerase chain reaction for visual colorimetric detection of amplicons.
abstract: We have developed a self-reporting polymerase chain reaction (PCR) system for visual colorimetric gene detection and distinction of single nucleotide polymorphisms (SNPs). Amplification is performed using target-specific primers modified with a 5'-end tail that is complementary to a G-quadruplex deoxyribozyme-forming sequence. At end-point, G-quadruplexes are forced to fold from PCR-generated duplex DNA and then are used to colorimetrically report the successful occurrence of PCR by assaying their peroxidase activity using a chromogenic substrate. Furthermore, primer design considerations for the G-quadruplex-generating PCR system have allowed us to visually distinguish SNPs associated with Mycobacterium tuberculosis drug resistance alleles.
authors.affiliation: Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Sanchita
authors.initials: S
authors.lastname: Bhadra
authors.forename: Vlad
authors.initials: V
authors.lastname: Codrea
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Ellington
mesh_terms: D000483:Alleles; D001426:Bacterial Proteins / Q000235:genetics; D002863:Chromogenic Compounds / Q000737:chemistry; D003124:Colorimetry; D004247:DNA / Q000032:analysis*; D017931:DNA Primers / Q000378:metabolism; D021881:DNA, Catalytic / Q000378:metabolism; D024881:Drug Resistance, Bacterial / Q000235:genetics; D054856:G-Quadruplexes; D009169:Mycobacterium tuberculosis / Q000235:genetics; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide
chemical_list: D001426:Bacterial Proteins; D002863:Chromogenic Compounds; D017931:DNA Primers; D021881:DNA, Catalytic; D004247:DNA
keywords: Colorimetric detection; G-quadruplex; Mycobacterium tuberculosis; Polymerase chain reaction; Single nucleotide polymorphism