Preparation method of calcium carbonate coated DNA-fluorescent dye composite nano gel dual anti-counterfeiting coating
A fluorescent dye and nanogel technology is applied in the field of preparation of calcium carbonate-coated DNA-fluorescent dye composite nanogel double anti-counterfeiting coatings, which can solve the problems of complicated and time-consuming operations, samples that cannot be reused, lack of clear anti-counterfeiting methods, and the like. High biocompatibility effect
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Embodiment 1
[0029] according to figure 1 The shown route prepares calcium carbonate-wrapped DNA-fluorescent dye composite nanogel light and dark double anti-counterfeiting coatings, and the specific steps are as follows:
[0030] Step 1: Collect the DNA sequence from the plasmid DNA, use the Primer Pemier 5.0 software to design primers, and intercept and amplify a DNA fragment with a length of 720 bp by PCR. The DNA amplification conditions are as follows: 50 μL system, including 25 μL Taq DNA polymerase mixture, 1 μL 1 mg / mL template DNA, 20 μmol / L upstream and downstream primers, 1 μL each and 22 μL deionized water; the amplification procedure refers to Figure 5 A: Pre-denaturation at 95°C for 10 minutes; denaturation at 95°C for 60 seconds, annealing at 60°C for 60 seconds, extension at 72°C for 60 seconds, repeat 25 cycles; extension at 72°C for 5 minutes to complete the extension of the amplified DNA sequence and terminate the reaction.
[0031]Step 2: Configure gelatin solution at...
Embodiment 2
[0039] according to figure 1 The route shown prepares calcium carbonate-wrapped DNA-fluorescent dye composite nanogel double anti-counterfeiting coating, and the specific steps are as follows:
[0040] Step 1: Collect DNA sequence from λ-DNA, use Primer Pemier 5.0 software to design primers and intercept and amplify a DNA fragment with a length of 200 bp by PCR. DNA amplification conditions are as follows: 50 μL system, including 25 μL TaqDNA polymerase mixture, 1 μL 1mg / mL template DNA, 20 μmol / L upstream and downstream primers 1 μL each and 22 μL deionized water; amplification program: 95 ° C pre-denaturation for 10 minutes Denaturation at 95°C for 60 seconds, annealing at 60°C for 60 seconds, extension at 72°C for 60 seconds, repeated for 25 cycles; extension at 72°C for 5 minutes to complete the extension of the amplified DNA sequence and terminate the reaction.
[0041] Step 2: Prepare a gelatin solution at a concentration of 10% (w / v), and dissolve it completely at 40°C...
Embodiment 3
[0049] according to figure 1 The shown route prepares calcium carbonate-wrapped DNA-fluorescent dye composite nanogel light and dark double anti-counterfeiting coatings, and the specific steps are as follows:
[0050] Step 1: Collect the DNA sequence from the plasmid DNA, use the Primer Pemier 5.0 software to design primers, and intercept and amplify a DNA fragment with a length of 350 bp by PCR. DNA amplification conditions are as follows: 50 μL system, including 25 μL Taq DNA polymerase mixture, 1 μL 1 mg / mL template DNA, 20 μmol / L upstream and downstream primers 1 μL and 22 μL deionized water; amplification program: 95 ° C pre-denaturation for 10 minutes Denaturation at 95°C for 60 seconds, annealing at 60°C for 60 seconds, extension at 72°C for 60 seconds, repeated for 25 cycles; extension at 72°C for 5 minutes to complete the extension of the amplified DNA sequence and terminate the reaction.
[0051] Step 2: Prepare a gelatin solution at a concentration of 5% (w / v), and...
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