Fluorescence quantitative PCR kit for detecting virus of aftosa and application
A foot-and-mouth disease virus and fluorescence quantitative technology are applied to a fluorescence quantitative PCR kit for detecting foot-and-mouth disease virus and its application field, and can solve the problem of inability to calculate the initial DNA or RNA copy number and the like
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Embodiment 1
[0030] The composition and preparation of the fluorescent quantitative PCR kit of embodiment 1 foot-and-mouth disease virus and its reaction conditions:
[0031] A). Kit composition:
[0032] a) The composition of the kit is as follows: (10 reactions)
[0033] RNA extraction solution A (4ml / tube) RNA extraction solution B (800μl / tube)
[0034] One-step amplification reaction solution (315μl / tube) Reverse transcription and hot start Taq DNA polymerase (10μl / tube)
[0035] Strong positive standard (100 times diluted PCR reaction tube (sterile, RNase and DNase free)
[0036] Release standard product 20μl / tube, four tubes in total) DEPC H 2 O (2ml / tube)
[0037] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0038] RNase inhibitor (400U / tube)
[0039] RNA extraction solution A is a product of Invitrogen Company, b) reverse transcriptase and hot start Taq DNA polymerase (2U / μl), c) one-step amplification (RT-PCR) reaction solution are all purchased f...
Embodiment 2
[0052] Embodiment 2 Fluorescence quantitative PCR kit detects the detection of different types of foot-and-mouth disease virus and compares it with plaque titration, a traditional method for quantitative determination of virus titer
[0053] A). Kit composition:
[0054] a) The composition of the kit is as follows: (10 reactions)
[0055] RNA extraction solution A (4ml / tube) RNA extraction solution B (800μl / tube)
[0056] One-step amplification reaction solution (315μl / tube) Reverse transcription and hot start Taq DNA polymerase (10μl / tube)
[0057] Strong positive standard (100 times diluted PCR reaction tube (sterile, RNase and DNase free)
[0058] Release standard product 20μl / tube, four tubes in total) DEPC H 2 O (2ml / tube)
[0059] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0060] RNase inhibitor (400U / tube)
[0061] RNA extraction solution A is a product of Invitrogen Company, b) reverse transcriptase and hot start Taq DNA polymerase (...
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