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Kit and DNA probe for detecting potato virus and preparation method of probe sequence (1)

A technology of DNA probes and viroids, applied in the field of preparation of kits for detecting potato viroids and DNA probes and probe sequences for detecting potato viroids, which can solve the problems of low detection sensitivity, complicated operation, and detection sensitivity Low-level problems, to achieve the effect of high sensitivity and simple operation

Inactive Publication Date: 2010-06-02
黑龙江省农业科学院植物脱毒苗木研究所
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Problems solved by technology

[0003] The present invention aims to solve the defects that existing potato viroid detection technology has low detection sensitivity, relatively complicated operation and requires professionals to carry out laboratory testing, and the existing DNA probes used for detecting potato viroids cause low detection sensitivity. Defects, but provide a kit for detecting potato viroids and a DNA probe for detecting potato viroids and a method for preparing the probe sequence ①

Method used

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  • Kit and DNA probe for detecting potato virus and preparation method of probe sequence (1)
  • Kit and DNA probe for detecting potato virus and preparation method of probe sequence (1)
  • Kit and DNA probe for detecting potato virus and preparation method of probe sequence (1)

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specific Embodiment approach 1

[0028] Specific embodiment one: the test kit that this embodiment detects potato viroid is by volume parts by 1 part of solution I, 1 part of solution VI, 1 part of solution VII, 1 part of solution VIII, 1 part of solution IX, 1 part of solution XI, 1 part of solution XII, 1 part of solution XIII and 1 part of solution XIV; wherein solution I is made of a mixed solution of sodium chloride and trisodium citrate, and the concentration of sodium chloride in solution I is 3mol / L, the concentration of trisodium citrate is 0.3mol / L, and its pH value is 7.0; the concentration of salmon sperm DNA in solution VI is 10mg / mL; solution VII is a mixture of solution V and solution IV according to the volume ratio of 101:1 Composed; every 10mL of solution V consists of 5mL of formamide, 3.5mL of 20% sodium dodecylsulfonate solution, 0.5mL of 0.5mL of 0.5mL of solution III and a balance of 5 mol of NaCl / L of aqueous solution; Na in solution II2 HPO 4 The concentration is 0.684mol / L, NaH 2 ...

specific Embodiment approach 2

[0030] Specific embodiment two: the DNA probe that this embodiment detects potato viroid is a double-body probe; The double-body probe has one of the following sequences:

[0031] ①、TCTAGATCCCTGAAGCGCTCCTCCGAGCCGCCTTTCTTTTTTCTTTTCT

[0032] GCTCAGGAGGTCAGGTGTGAACCACAGGAACCACGAGTTTAGTTCCGA

[0033] GGAACCAACTGCGGTTCCAAGGGCTAAACACCCTCGCCCCGAAGCAAA

[0034] GAAAGATAGAGAAAAAGCGGTTCTCGGGAGCTTCAGTTGTTTCTACCGG

[0035] GTAGTAGCCGAAGCGACAGCGCAAAGGGGGCGAGGGGTGGTCCTGCGG

[0036] GCGCGAGGAAGGACACCCGAAGAAAGGAAGGGTGAAAACCCTGTTTCG

[0037] GCGGGAATTACTCCTGTCGGCCGCTGGGCGCTCCCCACCGTCCTTATTGC

[0038] CAGTTCGCTCCAGGTTTCCCGGGGGATCCCTGAAGCGCTCCTCCGAGCC

[0039] GCCTTCTTTTTTCTTTTCTGCTCAGGAGGTCAGGTGTGAACCACAGGAAC

[0040] CACGAGTTTAGTTCCGAGGAACCAACTGCGGTTCCAAGGGCTAAACACC

[0041] CTCGCCCCGAAGCAAAGAAAGATAGAGAAAAAGCGGTTCTCGGGAGCT

[0042] TCAGTTGTTTCTACCGGGTAGTAGCCGAAGCGACAGCGCAAAGGGGGCG

[0043] AGGTGTGGTCCTGCGGGCGCGAGGAAAGACACCCGAAGAAAGGAAGG

[0044] GTGAAAACCCTGTTTCGGCGGGAATTACTCCTGTCGG...

specific Embodiment approach 4

[0048] Specific embodiment four: the DNA probe that the present embodiment detects potato viroid is double body probe; Double body probe has following sequence:

[0049] ①、TCTAGATCCCTGAAGCGCTCCTCCGAGCCGCCTTTCTTTTTTCTTTTCT

[0050] GCTCAGGAGGTCAGGTGTGAACCACAGGAACCACGAGTTTAGTTCC

[0051] GAGGAACCAACTGCGGTTCCAAGGGCTAAACACCCTCGCCCCGAAGCA

[0052] AAGAAAGATAGAGAAAAAGCGGTTCTCGGGAGCTTCAGTTGTTTCTACC

[0053] GGGTAGTAGCCGAAGCGACAGCGCAAAGGGGGCGAGGGGTGGTCCTGC

[0054] GGGCGCGAGGAAGGACACCCGAAGAAAGGAAGGGTGAAAACCCTGTT

[0055] TCGGCGGGAATTACTCCTGTCGGCCGCTGGGCGCTCCCCACCGTCCTTAT

[0056] TGCCAGTTCGCTCCAGGTTTCCCGGGGGATCCCTGAAGCGCTCCTCCGA

[0057] GCCGCCTTCTTTTTTTCTTTTCTGCTCAGGAGGTCAGGTGTGAACCACAGG

[0058] AACCACGAGTTTAGTTCCGAGGAACCAACTGCGGTTCCAAGGGCTAAAC

[0059] ACCCTCGCCCCGAAGCAAAGAAAGATAGAGAAAAAGCGGTTCTCGGGA

[0060] GCTTCAGTTGTTTCTACCGGGTAGTAGCCGAAGCGACAGCGCAAAGGGG

[0061] GCGAGGTGTGGTCCTGCGGGCGCGAGGAAAGACACCCGAAGAAAGGA

[0062] AGGGTGAAAACCCTGTTTCGGCGGGAATTACTCCTGTCGGCCGCTGGGC...

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Abstract

A kit and DNA probe for detecting potato viroid and a preparation method of probe sequence (1) relate to a kit for detecting viroid, a DNA probe for detection and a preparation method of probe sequence. The invention solves the defects that the existing potato viroid detection technology has low detection sensitivity and complicated operation and needs the professional to perform laboratory tests, and also solves the defect that the existing DNA probe used for detecting the potato viroid can cause low detection sensitivity. The kit of the invention comprises a solution I, a solution VI, a solution VII, a solution VIII, a solution IX, a solution XI, a solution XII, a solution XIII and a solution XIV. The DNA probe of the invention is a double probe. The kit of the invention has high sensitivity and easy operation, and the sensitivity of the DNA probe of the invention is 500 times of the sensitivity of the current simple probe.

Description

technical field [0001] The invention provides a kit for detecting viroids, a DNA probe for detection and a method for preparing the probe sequence. Background technique [0002] There are two main detection techniques for potato virus-like virus (PSTVd) in China: one is the R-PAGE method (reciprocating two-way polyacrylamide gel electrophoresis), which has low detection sensitivity and complex operation needs The other is the RT-PCR method, which has the problem of high detection cost. In addition, the existing DNA probes used in the detection of potato viroids are all monomer probes or fragment probes, and this type of DNA probes has also caused the defect of low detection sensitivity. Contents of the invention [0003] The present invention aims to solve the defects that existing potato viroid detection technology has low detection sensitivity, relatively complicated operation and requires professionals to carry out laboratory testing, and the existing DNA probes used f...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12P19/34C12R1/94
Inventor 吕典秋刘尚武邱彩玲王绍鹏宿飞飞李勇高云飞王文重张抒李学湛王亚洲
Owner 黑龙江省农业科学院植物脱毒苗木研究所
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