One-step RT-PCR (Reverse Transcription Polymerase Chain Reaction) detection reagent kit for Cucumber green mottle mosaic virus and detection method thereof
An RT-PCR and detection kit technology, applied in the field of one-step RT-PCR detection kits for cucumber green mottle mosaic virus, can solve the problems of low reliability, long detection period, complicated processing, etc. The effect of reducing detection cost and simple production method
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Example Embodiment
[0029] Example 1:
[0030] A one-step RT-PCR detection kit for cucumber green mottle mosaic virus, characterized in that the kit includes:
[0031] 1) A # The tube contains the upstream primers of Cucumber Green Mottle Mosaic Virus, the concentration is 10μmol / L, and the primer sequence is 5'-TGCTTCTTATGTTCCCGTCA-3';
[0032] 2) B # The tube contains the downstream primers of cucumber green mottle mosaic virus, the concentration is 10 μmol / L, and the primer sequence is 5’-GCCCATAGAAACTTCAACGTC-3’;
[0033] 3) C # Tube, containing PCR Buffer, the concentration is 10×;
[0034] 4)D # Tube with Mgcl inside 2 , The concentration is 25mmol / L;
[0035] 5)E # Tube with dNTPs inside, concentration of 10mmol / L;
[0036] 6)F # Tube with RNase inhibitor inside, the concentration is 40U / μL;
[0037] 7) G # Tube containing reverse transcriptase at a concentration of 5U / μL;
[0038] 8) H # The tube contains DNA polymerase at a concentration of 5U / μL;
[0039] 9) I # Tube containing positive control CK+ co...
Example Embodiment
[0043] Example 2:
[0044] The detection method of the cucumber green mottle mosaic virus one-step RT-PCR detection kit includes the following steps:
[0045] 1) Add A to each PCR tube according to the number of samples to be tested # Solution 2μL, B # Solution 2μL, C # Solution 5μL, D # 10μL, E # Solution 5μL, F # 1μL, G # 1μL, H # 1μL and K # 21μL of liquid, mix well, and leave after instant;
[0046] 2) RT-PCR reaction: Take 2μL each of the sample to be tested, the negative control and the positive control RNA template, add them to each PCR tube prepared in step (1), mix well, and perform the following RT-PCR reaction after instant separation: CDNA synthesis at 42°C for 30 minutes, followed by pre-denaturation at 94°C for 5 minutes, and then enter the following cycle: denaturation at 94°C for 30s, annealing at 54°C for 45s, extension at 72°C for 1 min, a total of 35 cycles; after the last cycle, extension at 72°C for 10 min;
[0047] 3) PCR amplification product electrophoresis det...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap