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Reference gene for fluorescence quantification of different tissues of Chinese yam and primers and application thereof

An internal reference gene and fluorescence quantitative technology, which is applied in the fields of application, genomics, genetic engineering, etc., can solve problems such as the stability of internal reference genes that have not been analyzed

Active Publication Date: 2021-03-30
YUNNAN AGRICULTURAL UNIVERSITY +1
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Problems solved by technology

[0003] There are few reports on internal reference genes of yam, only the cloning and expression of anthocyanidin synthase (ANS) gene, anthocyanin-related gene DaF3H and flavonol synthase DaFLS1 gene, among which Actin2 and Actin1 are used as internal reference genes Expression analysis, but the stability of reference genes has not been analyzed

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  • Reference gene for fluorescence quantification of different tissues of Chinese yam and primers and application thereof
  • Reference gene for fluorescence quantification of different tissues of Chinese yam and primers and application thereof
  • Reference gene for fluorescence quantification of different tissues of Chinese yam and primers and application thereof

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Embodiment Construction

[0025] The present invention will be further described in detail below in conjunction with the examples.

[0026] Those skilled in the art will understand that the following examples are only for illustrating the present invention and should not be considered as limiting the scope of the present invention. If no specific technique or condition is indicated in the examples, it shall be carried out according to the technique or condition described in the literature in this field or according to the product specification. The materials or equipment used are not indicated by the manufacturer, and they are all conventional products that can be obtained through purchase.

[0027] 1 Implementation materials

[0028] Using oxtail yam as the implementation material, select 1.5-3g / piece of complete zero seeds every other year to plant in the vegetable greenhouse or plant implementation platform of Yunnan Agricultural University, and first germinate and then plant and manage. Four diff...

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Abstract

The invention relates to a reference gene for fluorescence quantification of different tissues of Chinese yam and primers and application thereof, and belongs to the technical field of molecular biology of Chinese yam. The nucleotide sequence of the reference gene CKI-2 is as shown in SEQ ID NO. 1. The nucleotide sequence of a PCR amplification primer of the reference gene CKI-2 is as follows: anupstream primer is shown as SEQ ID NO.2; and the downstream primer is as shown in SEQ ID NO. 3. Protein kinase I, myb, F-box, actin gene, EF1, bHLH13, eIF1, grid heavy chain protein and other genes are selected as candidate genes of different tissues. The stability of candidate genes is evaluated through qRT-PCR in combination with RefFind, reference genes stably expressed in different tissues ofthe Chinese yams are selected out through screening, and a reference basis is provided for later development of research work such as gene function verification of growth and development and metabolicmechanisms of the Chinese yams.

Description

technical field [0001] The invention belongs to the technical field of molecular biology of yam yam, and specifically relates to an internal reference gene for fluorescence quantification of different tissues of yam, primers and applications thereof. Background technique [0002] One of the most conventional and effective methods for gene expression research, real-time fluorescent quantitative PCR (quantitative Real-time PCR, qRT-PCR) technology, by adding specific fluorescent groups to the PCR reaction system, can detect the products in the entire PCR reaction process. Changes are monitored in real time to achieve qualitative and quantitative expression analysis of target genes. However, qRT-PCR is affected by factors such as the quality of RNA, the quality of template cDNA, the specificity of primers, and the amplification rate of PCR. During the qRT-PCR reaction, it is necessary to select appropriate internal reference genes for calibration and standardization. At presen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12Q1/6809C12Q1/6895C12N15/11G16B20/00G16B30/00
CPCC12N9/12C12Q1/6809C12Q1/6895C12Y207/11001G16B20/00G16B30/00C12Q2600/158C12Q2600/166C12Q2531/113C12Q2563/107
Inventor 龙雯虹孙一丁唐文芳王仕玉段延碧徐升胜尹冬
Owner YUNNAN AGRICULTURAL UNIVERSITY
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