Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Sophora japonica L. transcriptome sequence-based and developed EST-SSR (expressed sequence tags-simple sequence repeats) marker primer and application thereof

A technology of transcriptome sequence and labeled primers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve problems affecting the innovative research and utilization of Chinese Sophora japonica germplasm resources, and achieve electrophoretic bands Clear, polymorphic rich, stable amplification results

Active Publication Date: 2018-08-17
SHANDONG FOREST SCI RES INST
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few studies on the molecular aspects of Sophora japonica, especially the analysis of genetic diversity of Sophora japonica is rarely reported, which seriously affects the innovative research and utilization of Sophora japonica germplasm resources
However, there is no report on the development of EST-SSR marker primers using the transcriptome sequence of Sophora japonica.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Sophora japonica L. transcriptome sequence-based and developed EST-SSR (expressed sequence tags-simple sequence repeats) marker primer and application thereof
  • Sophora japonica L. transcriptome sequence-based and developed EST-SSR (expressed sequence tags-simple sequence repeats) marker primer and application thereof
  • Sophora japonica L. transcriptome sequence-based and developed EST-SSR (expressed sequence tags-simple sequence repeats) marker primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1 Test materials and reagents

[0044] 1.1 Sophora japonica material

[0045] The experimental materials of this study are 233 germplasms of ancient Chinese pagoda tree (Table 1), mainly from Shandong Weifang, Tai'an, Dongying, Linyi, Heze, Yantai, Binzhou, Dezhou, Zibo, Jining, Rizhao, Liaocheng, Zaozhuang. old tree. All the germplasm of Chinese pagoda japonica ancient trees were collected from the original site in 2012 and then grafted and preserved in Yinmaquan Nursery and Zhangqiu Experimental Base of Shandong Academy of Forestry Sciences. The germplasm materials of Chinese pagoda tree ancient trees used in this study basically involve the ancient Chinese pagoda tree of various cities, counties and townships in Shandong Province.

[0046] Table 1 233 germplasms of Chinese pagoda japonica ancient trees tested

[0047]

[0048]

[0049]

[0050]

[0051]

[0052] 1.2 Primer acquisition

[0053] The high-throughput sequencing platform Illumina HiSeq ...

Embodiment 2

[0057] 2.1 Extraction of Genomic DNA from Sophora japonica

[0058] The genomic DNA of Sophora japonica was extracted using the improved CTAB method, and the young leaves of Sophora japonica were extracted as materials. And improve according to the characteristics of Chinese pagoda tree containing more phenols and protein.

[0059] 1. Preheat DNA extraction buffer 2×CTAB (100mM Tris-HCl (PH8.0), 1.4mM NaCl, 50MmEDTA (PH8.0), 2% CTAB) 700μL at 65℃, 20μL of β-mercaptoethanol (add now use), L, mix thoroughly.

[0060] 2. Weigh 0.5g of young Sophora japonica leaves, put them into a 2mL centrifuge tube, add two steel balls with a diameter of about 2mm, put them in a box filled with liquid nitrogen, freeze for about 15 seconds, and grind them with a retsch mixing grinder About 1.5min, until the leaves of Sophora japonica are in the form of fine powder, quickly add preheated L, and then put it into a 65°C water bath for about 30 minutes. During the water bath, pay attention to ...

Embodiment 3

[0098] 3.1 Detection of the concentration and purity of Genomic DNA of Sophora japonica

[0099] High-quality DNA is an important prerequisite to ensure the success of PCR amplification. The leaves of Sophora japonica contain secondary substances such as phenols and polysaccharides. These substances can inhibit the activity of Ex Taq enzyme and should be removed as much as possible during DNA extraction. In this study, PVP and β-mercaptoethanol were added to the DNA extraction liquid, and the modified CTAB method was used to extract the extracted DNA. After the UV spectrophotometer detected the extracted DNA, the results showed that the OD260 / OD280 value of the DNA was between 1.7 and 2.0 . The DNA was detected by 0.8% agarose gel electrophoresis, and the results were as follows: figure 1 As shown, it can be seen from the figure that the extracted DNA showed clear and complete bands after agarose gel electrophoresis without obvious tailing. This shows that the concentration ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the field of development and application of molecular marker technology and particularly relates to a sophora japonica L. EST-SSR marker primer group. The sophora japonica L.EST-SSR marker primer group is composed of primer pairs from No.1 to No.10. The sophora japonica L. EST-SSR marker primer group has the advantages of being stable in amplification, clear in electrophoretic bands, rich in polymorphism and the like and can be effectively applied to the research fields of sophora japonica L. such as planting resource genetic diversity analysis, structuring of high-density maps, variety purity and authenticity identification and molecular marker-assisted breeding.

Description

technical field [0001] The invention belongs to the field of development and application of molecular marker technology, and in particular relates to EST (expressed sequence tags, ESTs) microsatellite marker primer set (EST-SSR). Background technique [0002] Sophora japonica L., also known as Chinese pagoda tree, Chinese pagoda tree, pagoda tree, etc., belongs to the leguminous family Leguminosaesp. Sophora japonica L. is a deciduous tree with a height of 25 meters. It is native to the north of my country and has a profound cultivation history in China. As early as the Qin and Han Dynasties, there were already records about planting locust trees along the passageways. Chinese pagoda tree is a positive tree species, slightly shade-tolerant, not strict on soil requirements, well-developed root system, fast growth, high transplanting survival rate, strong adaptability, and also has the characteristics of anti-pollution, smoke resistance, wind resistance, etc., can be extremely...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11C12Q1/6806
CPCC12Q1/6806C12Q1/6895C12Q2600/156
Inventor 庞彩红李双云夏阳张明静付茵茵臧真荣刘盛芳杨勇亓玉昆李自峰王守国
Owner SHANDONG FOREST SCI RES INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com