Broccoli Ogu cytoplasm quick identification kit and detection method thereof

A detection method, broccoli technology, applied in the biological field, can solve the problems of long identification period, inability to accurately identify the specific type of sterile cytoplasm, time-consuming and labor-consuming operations, etc., and achieve the goal of shortening the identification period and rapid identification period Effect

Inactive Publication Date: 2011-05-25
WENZHOU VOCATIONAL COLLEGE OF SCI & TECH
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Problems solved by technology

[0006] Traditional CMS identification and classification methods such as tester lines, cytological markers, protein markers, isoenzyme identification, etc. have disadvantages such as long identification period, time-consuming and lab...

Method used

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  • Broccoli Ogu cytoplasm quick identification kit and detection method thereof
  • Broccoli Ogu cytoplasm quick identification kit and detection method thereof

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Embodiment Construction

[0043] Through the following examples and appended figure 1 The specific description of the present invention is only used to further illustrate the present invention, and cannot be interpreted as limiting the protection scope of the present invention. Technical engineers in this field can make some non-essential improvements and adjustments to the present invention according to the content of the above invention.

[0044] 1. Design and synthesis of specific primers

[0045] Primers were designed according to the gene orf138 related to Ogu CMS (Gene bank accession number: Z18896.1), and the sequence was synthesized in Huada Gene Company. The specific primers Ogu-A and Ogu-B are respectively located at 22bp-46bp and 394bp-413bp of the orf138 gene, and the length of the amplified specific fragment is 392bp.

[0046] 2. Assembling the kit

[0047] Preparation of Reagent A:

[0048]Centrifuge the tubes containing the primers (Ogu-A: 5’-TTGTCCACTTTTTGTCATAATCTCA-3’ and Ogu-B: 5’...

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Abstract

The invention discloses a broccoli Ogu cytoplasm quick identification kit and a detection method thereof. The kit comprises a reagent A and a reagent B, wherein the reagent A contains a PCR (Polymerase Chain Reaction) buffer solution, Taq polymerase, dNTP and an Oligo-dT primer, and the reagent B is standard DL-2000DNA Ladder. The detection method comprises the following steps of: DNA extraction of a sample; (1) genome DNA extraction of the sample; (2) PCR amplification reaction; (3) agarose gel electrophoresis detection by which whether the sample is Ogu cytoplasm or not is determined according to the existence/absence of 392bp specific amplification bands. By using the broccoli Ogu cytoplasm quick identification kit, the steps from DNA extraction to the obtaining of detection results can be finished only in several hours, which is very convenient and quick; and by the broccoli Ogu cytoplasm quick identification kit, screening and identifying can be finished in seedling stage, thereby greatly shortening identification cycle. The broccoli Ogu cytoplasm quick identification kit has very important value in broccoli breeding practices.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid identification kit for broccoli Ogu cytoplasm, which can detect broccoli Ogu cytoplasm and is suitable for rapid identification and molecular marker-assisted selection of the cytoplasm. Background technique [0002] Male sterility is a common natural phenomenon in plants, which means that due to various genetic or physiological reasons, the female organs of plants are normal, but the male organs are abnormal, unable to produce pollen or the pollen is aborted and cannot be pollinated normally, resulting in Self-crossing is not fruitful, only outcrossing is fruitful. Male sterility exists widely in the plant kingdom, and it has been found in many varieties or interspecific hybrids so far. [0003] Studies have shown that male sterility is mainly divided into two categories: nuclear male sterility (Genic male sterility, GMS) and cytoplasmic male sterility (abbreviated as CMS). ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 荆赞革唐征张小玲罗天宽刘庆叶珍朱世杨楼珏陈海英
Owner WENZHOU VOCATIONAL COLLEGE OF SCI & TECH
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