Method for detecting DNA of wheat stunt virus
A technology of wheat dwarf virus and DNA molecules, which is applied in biochemical equipment and methods, measurement/inspection of microorganisms, etc., can solve the problems of high price, high operator cost, unfavorable site, rapid analysis, etc., and achieve simple steps , strong sensitivity, easy to quickly detect and analyze the effect
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Embodiment 1
[0074] A method for detecting wheat dwarf virus DNA, the specific detection steps are as follows:
[0075] Step 1: Cut 100 mg of wheat diseased leaves, place the diseased leaves in a liquid mortar, add liquid nitrogen and grind them into powder; grind the virus-carrying plant tissue materials at room temperature until the juice fully flows out, and The plant tissue juice was obtained by centrifuging in a centrifuge, and heated at 93° C. for 30 seconds as the sample solution to be tested.
[0076] Step 2: Synthesize the DNA molecular machine and configure it into a solution with a concentration of 20nM,
[0077] Step 3: Take 4 μL of sample solution and 2 μL of DNA molecular machine solution, mix evenly by micro-oscillation to obtain a mixed solution,
[0078] Step 4: Add 10 μL buffer, 6 μL polymerase, 6 μL nickase, 6 μL Thioflavin T, 6 μL dNTP, 6 μL Mg 2+ Add to the mixed solution obtained in step 3 to obtain a reaction solution.
[0079] Wherein, the above-mentioned dNTP co...
Embodiment 2
[0098] A method for detecting wheat dwarf virus DNA, wherein the DNA molecular machine sequence is 5'-CCCCAAAACCCCAAAACCCCAAAACCCCTGCATCGACTCGAAGCACGAAGCTTG-3'.
[0099] The specific detection steps are as follows:
[0100] Step 1: Cut 100 mg of wheat diseased leaves, place the diseased leaves in a liquid mortar, add liquid nitrogen and grind them into powder; grind the virus-carrying plant tissue materials at room temperature until the juice fully flows out, and The plant tissue juice was obtained by centrifuging in a centrifuge, and heated at 93° C. for 30 seconds as the sample solution to be tested.
[0101] Step 2: Synthesize the DNA molecular machine and configure it into a solution with a concentration of 60nm,
[0102] Step 3: Take 8 μL of sample solution and 4 μL of DNA molecular machine solution, mix evenly by micro-oscillation, and obtain a mixed solution;
[0103] Step 4: Add 40 μL buffer, 10 μL polymerase, 10 μL nickase, 10 μL Thioflavin T, 10 μL dNTP, 10 μL Mg ...
Embodiment 3
[0113] A method for detecting wheat dwarf virus DNA, wherein the sequence of the DNA molecular machine is
[0114] 5'-CCCCAAAAACCCCAAAACCCCAAAACCCCTGCATCGACTCGAAGCACGAAGCTTG-3'.
[0115] The specific detection steps are as follows:
[0116] Step 1: Cut 100 mg of wheat diseased leaves, place the diseased leaves in a liquid mortar, add liquid nitrogen and grind them into powder; grind the virus-carrying plant tissue materials at room temperature until the juice fully flows out, and The plant tissue juice was obtained by centrifuging in a centrifuge, and heated at 93° C. for 30 seconds as the sample solution to be tested.
[0117] Step 2: Synthesize the DNA molecular machine and configure it into a solution with a concentration of 100nM,
[0118] Step 3: Take 4 μL of the sample solution and 4 μL of the DNA molecular machine solution, and mix evenly by micro-oscillation to obtain a mixed solution.
[0119] Step 4: Add 20 μL buffer, 6 μL polymerase, 7 μL nickase, 10 μL Thioflavi...
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