Visual detection method for ultrafast amplification of magnesium and zinc-cutting functional nucleic acids with universal partition
An amplification system and cleavage site technology, applied in the field of biosensors, can solve the problems of high price, complicated pretreatment, and long detection period.
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Embodiment 1
[0143] Example 1 Magnesium and zinc-cutting functional nucleic acid visual detection method for universal partition ultrafast amplification
[0144] 1. Experimental materials
[0145] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, magnesium chloride, zinc chloride, 2,2-azino-bis(3-ethane) Base-benzothiazole-6-sulfonic acid) diamine salt (ABTS), H 2 o 2 , 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), sodium hydroxide, and disodium hydrogen phosphate were all purchased from Thermo Scientific Life Technologies. All experimental water was from Milli-Q pure water system.
[0146] The sequence design is as follows (SEQ ID NO:1-11):
[0147]
[0148] Note: The ribozyme cuts the target product and the 3' end sequence of the amplified template is complementary;
[0149] The bold base sequence added to the end of the ribozyme substrate chain is to increase the Tm value of bin...
Embodiment 2
[0178] The specificity investigation of embodiment 2 detection method
[0179] According to the biosensor constructed in Example 1, 1 μM Mg 2+ , 500nM Zn 2+ , 10μM Ag + 、Cu 2+ 、Cd 2 + , Hg 2+ 、Cr 3+ , Fe 2+ , Ca 2+ , added to the system for detection, the results show that the established Mg 2+ , Zn 2+ The biosensor has good specificity ( Figure 8 ).
Embodiment 3
[0180] Embodiment 3 standard addition experiment
[0181] Get high-purity water and detect with the biosensor constructed in embodiment 1, Mg 2+ , Zn 2+ If there is no detection, the standard addition experiment is carried out on it, and the results obtained by continuous measurement are shown in Table 1 and Table 2.
[0182] Table 1 The results of the standard recovery experiment
[0183]
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