Screening Method of Microsatellite Markers in Seagrass Saltophila

A technology of microsatellite marker and screening method, which is applied in the field of molecular biology DNA marker technology and application, and can solve problems such as the limitation of genetic research of Salina philoxeroides

Inactive Publication Date: 2011-12-21
EAST CHINA NORMAL UNIV
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Problems solved by technology

However, due to the lack of microsatellite markers of Salina philodendron, the research on the genetics of S...

Method used

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  • Screening Method of Microsatellite Markers in Seagrass Saltophila
  • Screening Method of Microsatellite Markers in Seagrass Saltophila
  • Screening Method of Microsatellite Markers in Seagrass Saltophila

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Embodiment Construction

[0015] The screening of microsatellite sites and the determination of polymorphic markers of the present invention will be described in detail below through examples. The halophaga used in the experiment came from the seagrass beds of Hainan and Guangxi.

[0016] 1. Screening Microsatellite Sequences

[0017] Genomic DNA was extracted from leaf tissues of 12 halophila samples using a modified CTAB method. The specific operation of the improved CTAB method is as follows: (1) 0.05 g of the dry sample, which is dried on silica gel and wiped clean with 75% ethanol, is pulverized in a pulverizer for 40 seconds; (2) 1 ml of pre-cooled elution buffer (containing 0.02 g (3) centrifuge at 12000 rpm at 4°C for 15 minutes; (4) Discard the supernatant and add 780 μl 2×CTAB to extract buffer (containing 2.8 μl / ml of β-mercaptoethanol), in a water bath at 65°C for 2 hours, and shake up and down every 15 minutes; (5) add an equal volume of chloroform-isoamyl alcohol (volume ratio 24:1), sh...

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Abstract

The invention discloses a method for screening microsatellite markers of seagrass halophila. Firstly, the improved CTAB method is used to extract genomic DNA of the leaf tissue of halophila; , and the DNA fragments were sequenced, and DNA sequences with more than 5 base repeats (2-6) units were screened; then specific primers were designed at the flanking sequences at both ends of the microsatellite repeat sequence, and different geographical distribution groups The individual genome DNA of Salina philodendron was amplified by PCR to detect and optimize primers; finally, the fragment size of the PCR product was analyzed to determine the genotype of each individual, obtain the genetic polymorphism map of Salina philodendron, and determine the repeatability, Microsatellite markers with high stability and polymorphism. The invention can be used for the identification of germplasm resources, genetic diversity analysis, genetic map construction and other researches of the halophobia, and can be further used for the management, protection and restoration of the halophobia seagrass bed.

Description

technical field [0001] The invention belongs to the field of DNA marker technology and application of molecular biology, in particular to a screening method and application of microsatellite markers of seaweed Halophila ovalis (R.Br.) Hook. Background technique [0002] Halophila ovalis (R.Br.) Hook. is a perennial seagrass belonging to the genus Halophila ovalis of the family Hydrochardaceae. In the intertidal to subtidal soft mud and rough coral reefs (C.den Hartog et al., "Biology, Ecology and Conservation of Seagrasses", 2006, Chapter 2, pp. 25-50), hi my country Saltgrass is mainly distributed in the seagrass beds of Guangxi, Guangdong, Hainan and Hong Kong along the coast of South China (Huang Xiaoping, "Science Bulletin", 2006, Supplement II, pp. 136-152). However, in recent years, due to human activities and natural environment and other factors, seagrass beds have been declining. As human beings pay more attention to the ecological and economic functions of seagrass...

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
Inventor 徐娜娜刘敏陈小勇
Owner EAST CHINA NORMAL UNIV
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