Primer and method for rapid molecular detection of eelworm
A root-knot nematode and molecular detection technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA preparation, etc., can solve the problems of unstable and reliable test results, inability to identify larvae, long diagnosis time, etc. Guaranteed reliability, easy and fast operation, and strong practicability
Image
Examples
Embodiment 1
[0040] Southern (M.incognita), Java (M.javanica), peanut (M.arenaria) originating from China, Thailand, Iran, Australia, Netherlands, Belgium, Italy, Poland and the United States using primers MI-F and MI-R , northern (M.hapla) and root-knot nematode (M.enterolobii) 43 groups of genomic DNA carry out PCR amplification, the result only amplifies the product of 955bp on the root-knot nematode population (table 1; figure 1) . PCR reaction system 12.5μl, including 5ng nematode DNA, 0.4μM primers MI-F and MI-R, 0.2mM dNTPs, 1×ExTaq PCR buffer (including 2mM MgCl 2 ) and 1U ExTaq DNA polymerase (Takara Biotech, Dalian, China). The PCR conditions were: pre-denaturation at 94°C for 4min; 35 cycles of 94°C for 30s, 62°C for 30s and 72°C for 30s; and final extension at 72°C for 10min. The PCR instrument used was Eppendorf Mastercycler Personal (Eppendorf, Germany). Embodiment 2: Primer MJ-F / R is to the specific amplification of root-knot nematode javanica
Embodiment 2
[0041] Use primers MJ-F and MJ-R to pair the southern (M.incognita), Java (M.javanica), peanut (M.arenaria) originating from China, Thailand, Iran, Australia, the Netherlands, Belgium, Italy, Poland and the United States , northern (M.hapla) and root-knot nematode (M.enterolobii) 43 populations of genomic DNA carry out PCR amplification, the result only amplifies the product of 517bp on the root-knot nematode Java population (table 1; figure 2) . PCR reaction system 12.5μl, including 5ng nematode DNA, 0.4μM primers MJ-F and MJ-R, 0.2mM dNTPs, 1×ExTaq PCR buffer (including 2mM MgCl 2 ) and 1U ExTaq DNA polymerase (Takara Biotech, Dalian, China). The PCR conditions were: pre-denaturation at 94°C for 4min; 35 cycles of 94°C for 30s, 62°C for 30s and 72°C for 30s; and final extension at 72°C for 10min. The PCR instrument used was Eppendorf Mastercycler Personal (Eppendorf, Germany). Embodiment 3: The specific amplification of primer MI-F / MT-R to Southern, Java and peanut root-...
Embodiment 3
[0042] Southern (M.incognita), Java (M.javanica), peanut (M.arenaria) originating from China, Thailand, Iran, Australia, Netherlands, Belgium, Italy, Poland and the United States using primers MI-F and MT-R , northern (M.hapla) and root-knot nematode (M.enterolobii) 43 populations of genomic DNA were amplified by PCR, and as a result, a 779bp product was amplified on the southern, Java and peanut root-knot nematode populations, No amplified product (table 1; image 3) . PCR reaction system 12.5μl, including 5ng nematode DNA, 0.4μM primers MI-F and MT-R, 0.2mM dNTPs, 1×ExTaq PCR buffer (including 2mM MgCl 2 ) and 1U ExTaq DNA polymerase (Takara Biotech, Dalian, China). The PCR conditions were: pre-denaturation at 94°C for 4min; 35 cycles of 94°C for 30s, 62°C for 30s and 72°C for 30s; and final extension at 72°C for 10min. The PCR instrument used was Eppendorf Mastercycler Personal (Eppendorf, Germany). Example 4: Utilize primers MI-F / R, MJ-F / R and MI-F / MT-R to identify sin...
PUM
Login to View More Abstract
Description
Claims
Application Information
- IPC
- C12N15/10; C12P19/34; C12Q1/68
- Inventors
- 徐建华; 孟庆鹏