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Chinese cabbage EST-SSR marking primer and application thereof in variety identification

A technology for labeling primers and species identification, applied in the field of molecular biology, can solve problems such as development not reaching saturation

Active Publication Date: 2014-03-12
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the development of Chinese cabbage EST-SSR primers is far from being saturated

Method used

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  • Chinese cabbage EST-SSR marking primer and application thereof in variety identification
  • Chinese cabbage EST-SSR marking primer and application thereof in variety identification
  • Chinese cabbage EST-SSR marking primer and application thereof in variety identification

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Embodiment

[0038] 1. Development of Chinese cabbage EST-SSR marker primers

[0039] The EST sequences of Chinese cabbage used in the development of primers were obtained from GenBank (http: / / www.ncbi.nlm.nih.gov), and as of October 31, 2007, there were 147,217 EST sequences of Chinese cabbage (B.rapassp.pekinensis) . In this study, 147217 Chinese cabbage EST sequences were used to develop SSR primers.

[0040] The downloaded Chinese cabbage EST sequence was pre-processed according to the method of Ge Jia et al. (see "Journal of Agricultural Biotechnology", No. 04, 2005, "Analysis of Chinese Cabbage Expressed Sequence Tag SSR Marker"), including removing the EST sequence Existing vector sequence 5' end polyT and 3' end polyA sequence and sequences containing ambiguous bases, remove the sequence less than 100bp. Then, the SeqMan program (Lasergene software package, DNAStarInc.) was used to construct contigs (contigs) of the processed EST sequences. Use MISA software (http: / / www.pgrc.ipk...

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Abstract

The invention relates to a Chinese cabbage EST-SSR marking primer and application thereof in variety identification and belongs to the technical field of molecular biology. Sequences of Chinese cabbage EST-SSR marking primer BRE 131 and application of the marking primer in variety identification are provided. Amplification products of the EST-SSR primer presents parent complementary bands, size differences among bands are evident, hybrid and parental inbred line can be well distinguished, and different varieties can be identified. Amplification products of individuals in a same variety have consistent bands.

Description

[0001] This case is a divisional application with an application date of July 26, 2010 and an application number of 201010236157.6 technical field [0002] The invention relates to a Chinese cabbage EST-SSR marker primer and its application in variety identification, belonging to the technical field of molecular biology. Background technique [0003] Chinese cabbage (Brassicarapa L.ssp.pekinensis) is a Brassicaceae plant, and it is an important vegetable in my country and East Asian countries. Variety differences directly affect the production and eating quality of Chinese cabbage, so the identification of Chinese cabbage varieties and purity identification are very important. The most reliable traditional testing method for seed purity is the GOT (grow-out test) method, which uses field planting to identify hybrids based on morphological characteristics. The biggest disadvantage of the GOT method is that it is time-consuming and labor-intensive, and it is easily affected b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 高建伟刘立锋周新成李化银李利斌张庶
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI