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Method for detecting lily seed virus

A detection method, lily technology, applied in the detection of LSV and LMoV, lily bulb virus CMV field, can solve the problems of RNA take away, low quality of bulb total RNA extraction, RNA yield reduction, etc., to achieve easy distinction, total Inexpensive and reproducible results

Inactive Publication Date: 2010-02-17
BEIJING FORESTRY UNIVERSITY
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AI Technical Summary

Benefits of technology

This patented technology describes different methods for detecting viruses by create DNA fragments containing these genes together or separately. These techniques involve modifying certain parts (principle called nucleotide sequences), making them more compatible than others when used alone but still able to produce enough copies during replication without compromising their ability to function properly. By combining this modified nucleic acid into another form like plasmid before being introduced into cells, it becomes possible to make better use of genetic resources due to greater efficiency compared to existing systems. Additionally, there has been research on improving the accuracy of measuring small amounts of biological materials based upon enzymatic activity. Overall, this new technique allows us to efficiently study large numbers of samples and identify unknown agents quickly and accurately.

Problems solved by technology

Technological Problem addressed by this patented technical problem is how to efficiently isolate specific types of nucleic acids that can cause diseases like cytomegalovirus disease (CMD).

Method used

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  • Method for detecting lily seed virus
  • Method for detecting lily seed virus
  • Method for detecting lily seed virus

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Embodiment 2 3

[0059] The optimization of embodiment 2 triple compound RT-PCR effective system

[0060] In order to optimize the RT-MPCR system of the present invention, based on the RT-MPCR system established above, the components and concentrations in the RT system and the components and concentrations in the PCR system were studied. The basic principles of optimizing the PCR system are: obtaining clear and stable target-specific products; low cost; short reaction time, etc.

[0061] For the main component of RT, dNTPs, three pairs of primers, RNasin, M-MLV, and viral RNA concentrations, 6 groups of gradient treatments were set up. The main reaction components of PCR dNTPs, Mg 2+ , three pairs of primers, TaqDNA polymerase, template cDNA concentration and amplification time were each set to 6 groups of gradient treatments. The negative control uses the corresponding concentration in the established effective system.

[0062] (1) Effect of M-MLV reverse transcriptase concentration on RT-...

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Abstract

The invention provides a method for detecting lily seed virus including the detection of three virus of CMV, LSV and LMoV. The method comprises the following steps: (1) designing three pairs of virusprimers with good compatibility; (2) adopting improved Trizol and CTAB method to extract seedball squama RNA; and (3) using RT-MPCR technique to detect virus. The method for detecting lily seed virusis stable and fast, has good repetitiveness and low cost, and can be applied to detect the CMV, LSV and LMoV virus of the lily seedball.

Description

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Claims

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Application Information

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Owner BEIJING FORESTRY UNIVERSITY
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