Coupling an universal primer to SBE combined spectral codification strategy for single nucleotide polymorphism analysis.
We previously reported a strategy that combines Förster resonance energy transfer (FRET) based spectral codification with a single base extension (SBE) reaction for single nucleotide sequence discrimination in solution. This strategy is capable of unequivocally detect the allele variants present in solution. To extend the use of this tool to any locus of interest, it would be required the development of an universal approach capable of combining a sequence specific SBE primer to an universal sequence labeled and optimized for spectral codification. Here, we extend this concept to a general strategy by means of a labeled universal oligonucleotide primer (donor), a sequence specific primer that allows for incorporation of the complementary acceptor labeled ddNTP, which allows discrimination the allele variant in the sample via the unambiguous FRET signature of the donor/acceptor pair.
CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal; REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
Mílton
M
Cordeiro
Letícia
L
Giestas
João C
JC
Lima
Pedro
P
Baptista
D017931:DNA Primers / Q000235:genetics*; D031541:Fluorescence Resonance Energy Transfer; D008954:Models, Biological; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*
D017931:DNA Primers
FRET; SBE; SNP; Spectral codification; Universal SBE primer