Bovine parainfluenza type-3 virus detecting fluorescence quantitative PCR kit and application thereof
A type 3 virus, fluorescence quantitative technology, applied in fluorescence/phosphorescence, microbe-based methods, microbe assay/inspection, etc., can solve the problem of inability to calculate the initial DNA or RNA copy number
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Embodiment 1
[0030] The fluorescent quantitative PCR kit composition and reaction condition thereof of embodiment 1 bovine parainfluenza type 3 virus
[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] Reverse transcription reaction solution (90μl / tube) Reverse transcriptase (10μl / tube)
[0035] RNase inhibitor (400U / tube)
[0036] Strong Positive Standard (100-fold diluted standard 20μl / tube, four tubes in total)
[0037] PCR reaction tube (sterile, RNase and DNase free) DEPC H 2 O (2ml / tube)
[0038] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0039] PCR amplification reaction solution (220μl / tube) Hot start Taq DNA polymerase (2.75μl / tube)
[0040] (RNA extraction solution A is a product of Invitrogen Company. b) reverse transcription reaction solution, d) reverse transcriptase (purchased from Promega Company, g)...
Embodiment 2
[0060] Example 2: Application of Bovine Parainfluenza Type 3 Virus Fluorescent Quantitative PCR Kit in the Epidemiological Investigation of Cows Infected with Bovine Parainfluenza Type 3 Virus
[0061] A). Kit composition:
[0062] a) The composition of the kit is as follows: (10 reactions)
[0063] RNA extraction solution A (5ml / tube) RNA extraction solution B (5ml / tube)
[0064] RNA extraction solution C (10ml / tube)
[0065] Filter column A and adsorption column B (sterile, RNase and DNase free)
[0066] Reverse transcription reaction solution (90μl / tube) Reverse transcriptase (10μl / tube)
[0067] RNase inhibitor (400U / tube)
[0068] Strong Positive Standard (100-fold diluted standard 20μl / tube, four tubes in total)
[0069] PCR reaction tube (sterile, RNase and DNase free) DEPC H 2 O (2ml / tube)
[0070] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0071] PCR amplification reaction solution (220μl / tube) Hot start Taq DNA polymerase (2.75μl...
Embodiment 3
[0091] Example 3: Application of Bovine Parainfluenza Type 3 Fluorescent Quantitative PCR Kit in Detection of Commercially Available Bovine Serum
[0092] A). Kit composition:
[0093] a) The composition of the kit is as follows: (10 reactions)
[0094] RNA extraction solution A (4ml / tube) RNA extraction solution B (800μl / tube)
[0095] Reverse transcription reaction solution (90μl / tube) Reverse transcriptase (10μl / tube)
[0096] RNase inhibitor (400U / tube)
[0097] Strong Positive Standard (100-fold diluted standard 20μl / tube, four tubes in total)
[0098] PCR reaction tube (sterile, RNase and DNase free) DEPC H 2 O (2ml / tube)
[0099] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0100] PCR amplification reaction solution (220μl / tube) Hot start Taq DNA polymerase (2.75μl / tube)
[0101] (RNA extraction solution A is a product of Invitrogen Company. b) reverse transcription reaction solution, d) reverse transcriptase was purchased from Promega...
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