Genotyping detection method of drug resistance to Sanmate of Fusarium graminearum
A technology of Fusarium graminearum and typing method, which is applied in the field of genotyping and detection of Fusarium graminearum resistance to carbendazim, which can solve the problem of time-consuming, heavy workload, and inability to predict drug-resistant diseases in the early stage Pop and other issues
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0100] Embodiment 1, detect the genotype strain (position 167) of point mutation in the 167th codon
[0101] Extract bacterial strain ZF-43, ZF-21, ZF-52, R9 genomic DNA, use the specific primer amplification pair that amplifies Fusarium graminearum β2-tubulin gene to comprise 167 codon DNA fragments:
[0102] F-out167 5′GACCACCTTCAGGGTTTCCAGCTGACGC 3′
[0103] R-out167 5′GATCGGCGTACGAAGGATCGGCGATCTT 3′
[0104] F-in167T 5′GTTCCCCGATCGCATGATGGCCACTTT 3′
[0105] R-in167A 5′GAAACCTTGGGCGAGGGCATAACGGGAT 3′
[0106] 25 μl PCR system contains 2 μl DNA template, 2.5 μl 10× buffer, 2 μl dNTP (2.5 mM each), 1.5 μl MgCl2 (25 mM), 0.2 μl each of F-out167 / R-out167 (10 μM), F-in167T (10 μM) 0.5 μl, 0.8 μl of R-in167A (10 μM), 1 U of TaqDNA polymerase (Dalian Biotech, Takara). The reaction was carried out on PTC-200 (Bio-rad, Inc.), and the reaction conditions were: 94°C for 5 min; 35 cycles of 94°C for 15 s, 66°C for 30 s, and 72°C for 30 s; 72°C for 5 min; and stored at 4°C. Get 8 ...
Embodiment 2
[0108] Embodiment 2, detection of the genotype strain (position 198) in which a point mutation occurs in the 198th codon
[0109] Extract bacterial strain ZF43, ZF-21, J2, ZF43-6 genomic DNA, use the specific primer amplification pair that amplifies Fusarium graminearum β2-tubulin gene to comprise 198 codon DNA fragments:
[0110] F-out198 5′AGCTCGTTGAGGAAGCCATTGATGTT 3′
[0111] R-out198 5′CATGTTAACAGCGAGCTTTCGCAGAT 3′
[0112] F-in198A 5′GAACCAGCTCGTCGAGAACTCTGCCA 3′
[0113] R-in198G 5′GAGCCTCGTTATCGATACAGAAGGTATC 3′
[0114] 25 μl PCR system contains 2 μl DNA template, 2.5 μl 10× buffer, 2 μl dNTP (2.5 mM each), 1.5 μl MgCl2 (25 mM), 0.4 μl each of F-out198 / R-out198 (10 μM), F-in198A (10 μM) 0.15 μl, R-in198G 0.4 μl, TaqDNA polymerase 1U (Dalian Takara, Takara). The reaction was carried out on PTC-200 (Bio-rad, Inc.), and the reaction conditions were: 94°C for 5 minutes; 35 cycles of 94°C for 15s, 65°C for 30s, and 72°C for 30s; 72°C for 5 minutes; and stored at 4°C. ...
Embodiment 3
[0116]Embodiment 3, detection of the genotype strain (position 200) in which a point mutation occurs in the 200th codon
[0117] Extract bacterial strain ZF43, ZF-21, NT-7, t1 genomic DNA, amplify the DNA fragment specific primer pair that contains 200 codons with amplifying Fusarium graminearum β2-tubulin gene:
[0118] F-out200 5′ACAACTGGGCCAAGGGTCATTACACC 3′
[0119] R-out200 5′AAGTCGGGGGAACGGAATCATGTTAAC 3′
[0120] F-in200T 5′CCAGCTCGTCGAGAACTCTGACGAGACTTT 3′
[0121] R-in200A 5′TCGTACAGAGCCTCGTTATCGATACGGT 3′
[0122] 25μl PCR system contains 2μl DNA template, 2.5μl 10×buffer, 2μl dNTP (2.5mM each), 1.5μl MgCl2 (25mM), 0.8μl each of F-out200 / R-out200 (10μM), F-in200T / R- 0.5 μl each of in200A (10 μM), and 1 U of TaqDNA polymerase (Dalian Biotech, Takara). The reaction was carried out on PTC-200 (Bio-rad, Inc.), and the reaction conditions were: 94°C for 5 min; 35 cycles of 94°C for 15 s, 55°C for 30 s, and 72°C for 30 s; 72°C for 5 min; and stored at 4°C. Get 8 μ l o...
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, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com