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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.

SUBMITTER: Bhadra S

PROVIDER: S-EPMC3893031 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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G-quadruplex-generating polymerase chain reaction for visual colorimetric detection of amplicons.

Bhadra Sanchita S Codrea Vlad V Ellington Andrew D AD

Analytical biochemistry 20131014

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 the ...[more]

PMID: 24135653

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