Universal nucleic acid detection kit and method for human adenoviruses
A detection kit, human adenovirus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/testing, etc., can solve the problems of insensitivity of clinical specimens, susceptibility to bacterial and fungal contamination, etc. Achieve the effect of mature detection method, good specificity and reliable experimental basis
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Embodiment 1
[0022] A human adenovirus universal nucleic acid detection kit, which includes an extraction solution for extracting human adenovirus DNA, a PCR reaction solution, and the reaction solution includes 50 μL of 13 μM Taq DNA polymerase, 100 μL of 150 μM dNTP, 200 μL of SyBr dye, and 15 μM of uracil glycosyl Lyase 50 μL, human adenovirus positive control substance, negative control substance;
[0023] The primer sequences used in the DNA amplification reaction in the PCR reaction solution are as follows:
[0024] P1: 5'-ACATCGCCGGACAGGATGCTTCGGAGTACC-3';
[0025] P2: 3'-CACCCGCTGTTGTCTCACGACCTATACCGG-5'.
[0026] The PCR reaction system is 20 μL, including 15 μL of the PCR reaction solution, 2 μL of the primers and 3 μL of sample DNA;
[0027] The PCR reaction cycle parameters are:
[0028] 94°C, 2min; enter the cycle stage: denaturation at 94°C for 15s, annealing at 50°C for 1min, extension at 72°C for 20s, same as 35 cycles; stop at 10°C.
Embodiment 2
[0030] A human adenovirus universal nucleic acid detection kit, which includes an extraction solution for extracting human adenovirus DNA, a PCR reaction solution, and the reaction solution includes 50 μL of 13 μM Taq DNA polymerase, 100 μL of 150 μM dNTP, 200 μL of SyBr dye, and 15 μM of uracil glycosyl Lyase 50 μL, human adenovirus positive control substance, negative control substance;
[0031] The primer sequences used in the DNA amplification reaction in the PCR reaction solution are as follows:
[0032] P1: 5'-ACATCGCCGGACAGGATGCTTCGGAGTACC-3';
[0033] P2: 3'-CACCCGCTGTTGTCTCACGACCTATACCGG-5'.
[0034] The PCR reaction system is 20 μL, including 15 μL of the PCR reaction solution, 2 μL of the primers and 3 μL of sample DNA;
[0035] The PCR reaction cycle parameters are:
[0036] 94°C, 2min; enter the cycle stage: denaturation at 94°C for 15s, annealing at 55°C for 1min, extension at 72°C for 20s, same as 35 cycles; stop at 10°C.
Embodiment 3
[0038] A human adenovirus universal nucleic acid detection kit, which includes an extraction solution for extracting human adenovirus DNA, a PCR reaction solution, and the reaction solution includes 50 μL of 13 μM Taq DNA polymerase, 100 μL of 150 μM dNTP, 200 μL of SyBr dye, and 15 μM of uracil glycosyl Lyase 50 μL, human adenovirus positive control substance, negative control substance;
[0039] The primer sequences used in the DNA amplification reaction in the PCR reaction solution are as follows:
[0040] P1: 5'-ACATCGCCGGACAGGATGCTTCGGAGTACC-3';
[0041] P2: 3'-CACCCGCTGTTGTCTCACGACCTATACCGG-5'.
[0042] The PCR reaction system is 20 μL, including 15 μL of the PCR reaction solution, 2 μL of the primers and 3 μL of sample DNA;
[0043] The PCR reaction cycle parameters are:
[0044] 94°C, 2min; enter the cycle stage: denature at 94°C for 15s, anneal at 60°C for 1min, extend at 72°C for 20s, same as 35 cycles; stop at 10°C.
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