Rabies virus detecting fluorescence quantitative PCR kit and application thereof
A fluorescent quantitative and rabies virus technology, applied in the direction of fluorescence/phosphorescence, microbe-based methods, microbiological measurement/inspection, etc., can solve the problem of not being able to calculate the starting DNA or RNA copy number
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Embodiment 1
[0029] The fluorescent quantitative PCR kit composition of embodiment 1 rabies virus and reaction condition thereof
[0030] A). Kit composition:
[0031] a) The composition of the kit is as follows: (10 reactions)
[0032] RNA extraction solution A (4ml / tube) RNA extraction solution B (800μl / tube)
[0033] Reverse transcription reaction solution (90μl / tube) Reverse transcriptase (10μl / tube)
[0034] RNase inhibitor (400U / tube)
[0035] Strong Positive Standard (100-fold diluted standard 20μl / tube, four tubes in total)
[0036] PCR reaction tube (sterile, RNase and DNase free) DEPC H 2 O (2ml / tube)
[0037] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0038] Fluorescent quantitative PCR reaction solution (125μl / tube)
[0039] (RNA extraction solution A is a product of Invitrogen Company. b) reverse transcription reaction solution, d) reverse transcriptase was purchased from Promega Company, g) fluorescent quantitative PCR reaction solution an...
Embodiment 2
[0059] Example 2: Application of rabies virus fluorescent quantitative PCR kit in detection of rabies virus infection in cows
[0060] A). Kit composition:
[0061] a) The composition of the kit is as follows: (10 reactions)
[0062] RNA extraction solution A (5ml / tube) RNA extraction solution B (5ml / tube)
[0063] RNA extraction solution C (10ml / tube)
[0064] Filter column A and adsorption column B (sterile, RNase and DNase free)
[0065] Reverse transcription reaction solution (90μl / tube) Reverse transcriptase (10μl / tube)
[0066] RNase inhibitor (400U / tube)
[0067] Strong Positive Standard (100-fold diluted standard 20μl / tube, four tubes in total)
[0068] PCR reaction tube (sterile, RNase and DNase free) DEPC H 2 O (2ml / tube)
[0069] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0070] Fluorescent quantitative PCR reaction solution (125μl / tube)
[0071] (RNA extraction solution A, B, C and filter column adsorption column are products ...
Embodiment 3
[0090] Example 3: Application of rabies virus fluorescent quantitative PCR kit in detection of commercially available bovine serum
[0091] A). Kit composition:
[0092] a) The composition of the kit is as follows: (10 reactions)
[0093] RNA extraction solution A (4ml / tube) RNA extraction solution B (800μl / tube)
[0094] Reverse transcription reaction solution (90μl / tube) Reverse transcriptase (10μl / tube)
[0095] RNase inhibitor (400U / tube)
[0096] Strong Positive Standard (100-fold diluted standard 20μl / tube, four tubes in total)
[0097] PCR reaction tube (sterile, RNase and DNase free) DEPC H 2 O (2ml / tube)
[0098] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)
[0099] Fluorescent quantitative PCR reaction solution (125μl / tube)
[0100] (RNA extraction solution A is a product of Invitrogen Company. b) reverse transcription reaction solution, d) reverse transcriptase was purchased from Promega Company, g) fluorescent quantitative PCR reactio...
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