Part:BBa_K4612141
code1_F
code1_F(BBa_K4612141) is an 18-nucleotide single-stranded DNA sequence with the fluorescent moiety FAM modified at the 3' end. Together with the corresponding quenching probe code1_Q(BBa_K4612143) it forms a set of reporter probes. This set of reporter probes is used to convert a target signal with a positive weight number into a fluorescent signal after summation. In the absence of the target nucleic acid, the reporter probes bring the distance between the fluorescent molecules and the quenching molecules closer due to the formation of a stable double-stranded structure, resulting in fluorescence resonance energy transfer (FRET) occurring and fluorescence quenching; when the target is added, the weighted strand of the polymerase-mediated strand displacement hybridizes with the long DNA strand in the double-stranded probes through the toehold-mediated strand displacement reaction, resulting in the formation of a thermodynamically more stable double-stranded structure The DNA short chain is also competed down to achieve fluorescence recovery. Since different target DNA sequences have different weights in the previous step, different numbers of weighted strands will be released during DNA polymerization, and therefore different intensities of fluorescence signals will be released in this step of the reaction.
After the fluorescence curve is stable, the fluorescence intensity is linearly related with the concentration of the weight molecules, proving that the fluorescent reporter probe can produce a linear response to different concentrations of the weight molecules, which can realize the quantitative conversion of the weight molecular signal to the fluorescence intensity signal.
Sequence and Features
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