Part:BBa_K4612137

code_2

code_2(BBa_K4612137) is a single-stranded DNA sequence with 23 nucleotides, and is a universal weighted molecular with a negative weight number. The weighted molecular has its partial complementary sequence positioned after the target (5' to 3') in an amount determined by the weight number of the target. The 18 nt at the 5' end of the weighted molecular is paired with the weighted probe, and the last 5 nt is not paired with the weighted probe. The presence of this free 5 nt sequence prevents the DNA polymerase from combining with the weighted probe between the weighted molecular and initiating a polymerase-mediated strand displacement reaction, resulting in the number of weighted strands displaced, released and less than the corresponding weights of the target.



This part is the weight molecular in the figure, which binds to the weighted probe in the same number as the number of weights of the target (e.g., -4 in the figure), and is all displaced in a polymerase-mediated DNA strand displacement reaction, followed by a toe-mediated strand displacement reaction, which converts the signal of the number of weight molecules into a fluorescent signal.

1.After the fluorescence curve was stabilized, the fluorescence intensity was linearly correlated with the concentration of weight molecules, which proved that the fluorescent reporter probe could produce linear responses to different concentrations of weight molecules, and could realize the quantitative conversion of weight molecule signals to fluorescence intensity signals, and demonstrated the reliability of the weight molecules to be converted into fluorescence signals.

2.As shown, after stabilization of the fluorescence signal, the fluorescence intensity of the solution depends linearly on the target weight and the target concentration, demonstrating the ability to release the corresponding weight number of weight molecules by the polymerase-mediated chain substitution reaction.





3.The results showed that after the reaction, the fluorescence intensity of the mixed targets and the sum of the weight numbers were linearly related, and the individual targets did not interfere with each other in the binding of the weighted probes, and the weighted signal sum could be realized, proving the reliability of the weighted molecules in signal sum. Sequence and Features BBa_K4612137 SequenceAndFeatures