Feature sequences, tag primer and identification method of carya illinoensis variety Van Deman

A thin-shell hickory nut and labeling primer technology, applied in biochemical equipment and methods, microbial measurement/inspection, recombinant DNA technology, etc., can solve the problems of low yield, instability, difficult parent selection and rational allocation, etc., to achieve The effect of simple method and improved reliability

Active Publication Date: 2018-01-09
ZHEJIANG FORESTRY ACAD
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Problems solved by technology

[0003] However, the main problem faced by the existing pecan production areas in my country is low yield and instability. There are many reasons for this phenomenon, one of which is the lack of clear genetic relationship analysis of the existing introduced varieties Coupled with the confusion in the naming of some hybrid offspring, it is difficult to carry out effective parent selection and rational allocation, and it is not convenient for variety identification, promotion,

Method used

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  • Feature sequences, tag primer and identification method of carya illinoensis variety Van Deman
  • Feature sequences, tag primer and identification method of carya illinoensis variety Van Deman
  • Feature sequences, tag primer and identification method of carya illinoensis variety Van Deman

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Experimental program
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Embodiment 1

[0038] (1) Extraction of Genomic DNA of Thin-shelled Hickory Nut Variety:

[0039]Take 0.1g of the young leaves of the pecan variety to be tested, add liquid nitrogen and grind them thoroughly, and use the SDS-CTAB method to extract the genomic DNA. things. The crude DNA was purified by Magabio Nucleic Acid Purification Kit (Bioer, Hangzhou, China), and detected by 1.5% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (GeneQuant Pro, GE Healthcare). Integrity, Purity and Concentration. DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.

[0040] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0041] The upstream primer: 5'-ATGCAAGACGCTTGTGGAGA-3'; and the downstream primer: 5'-CCGACCCGACCCGATAAAAT-3' were synthesized by Shanghai Bioengineering Technology Co., Ltd.

[0042] (4) PCR amplification:

[0043] Composition of PCR reaction sol...

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Abstract

The invention relates to feature sequences and a molecular specificity tag primer of a pair of high-specificity feature sequences of carya illinoensis variety Van Deman, a molecular specificity tag primer, and a method capable of being used for fast identifying the carya illinoensis variety Van Deman. The molecular specificity tag primer has the sequence that a forward primer is 5'-ATGCAAGACGCTTGTGGAGA-3', and a reverse primer is 5'-CCGACCCGACCCGATAAAAT-3'. The molecular specificity tag primer provided by the invention can be used for performing fast early-stage identification on the carya illinoensis variety Van Deman; the method is simple, fast and accurate, and belongs to an irreplaceable molecular technique for distinguishing the carya illinoensis variety through appearance features.

Description

(1) Technical field [0001] The invention relates to a characteristic sequence of the pecan variety Van Deman, a molecular specific marker primer and a method for quickly identifying the pecan variety Van Deman by using the molecular specific marker primer. (2) Background technology [0002] Shelled Pecans (Carya (Wangenh.) K.Koch) is the most economically valuable tree species in the Juglandaceae Hickory genus. Carya carya is a typical outcrossing plant. According to the existing production practice, the variety configuration of carya carya is one of the main factors affecting its yield. The U.S. is one of the origins of pecans and its central production area. Over the years, it has selected and named about 1,000 cultivars. It has been more than 100 years since the introduction of hickory nuts in my country, and there are dozens of varieties commonly used in production at present. Years of production practice have proved that the vast subtropical regions of my country are...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 朱汤军彭华正金群英叶华琳
Owner ZHEJIANG FORESTRY ACAD
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