Unlock instant, AI-driven research and patent intelligence for your innovation.

Primer pair for Lilium EST (Expressed Sequence Tag) - SSR (Simple Sequence Repeat) detection

A primer pair, the technology of Lilium, applied in the field of molecular biology DNA marker technology and application

Inactive Publication Date: 2013-07-31
ZHEJIANG UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The number of molecular markers affects the accuracy of genetic diversity analysis of germplasm resources and the accuracy of genetic map construction. The number of existing molecular markers in Lilium is far from meeting the needs of molecular biology research in Lilium

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
  • Primer pair for Lilium EST (Expressed Sequence Tag) - SSR (Simple Sequence Repeat) detection
  • Primer pair for Lilium EST (Expressed Sequence Tag) - SSR (Simple Sequence Repeat) detection
  • Primer pair for Lilium EST (Expressed Sequence Tag) - SSR (Simple Sequence Repeat) detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0023] In the present invention, the specific method of developing the Lilium SSR marker with the EST sequence is:

[0024] 1. DNA extraction

[0025] (1) Prepare DNA extraction buffer: 2% CTAB, 0.1M Tris, 20mM EDTA, 1.4M NaCl, pH8.0 and DNA dissolution buffer TE: 10 Mm Tris; 1 Mm EDTA; PH=8.0.

[0026] Table 2 Different genotypes of lilies used in the realization of the present invention

[0027]

[0028] (2) The above-mentioned lily materials are processed as follows:

[0029] A. Quickly weigh about 0.2 g of young lily leaves stored in a -80°C refrigerator with a balance, and grind them in a mortar with liquid nitrogen. Transfer the powder into a 10 ml centrifuge tube containing 4 ml of CTAB solution and 80 μl of β-mercaptoethanol (preheated at 65°C), in a water bath at 65°C for 1 h; add 4 ml of chloroform / isoamyl alcohol (24:1), Mix well, centrifuge at 12,000 rpm for 10 min, take the supernatant and add 4 ml chloroform / isoamyl alcohol (24:1) again, mix well, and centr...

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 relates to the fields of molecular biology DNA (deoxyribonucleic acid) marking technology and application, and aims to provide four lilium (Expressed Sequence Tag) - SSR (Simple Sequence Repeat) markers and the application thereof. The four markers are respectively a group of primer pair of which the sequence is disclosed in SEQ ID NO: 1-8. The four molecular markers disclosed by the invention have polymorphism in 26 lily seeds or varieties, the genetic diversity analysis carried out in the 26 lily seeds or varieties is identical with the common sense of lilium plant classification (figure 1), the EST-SSR markers are stably existing new markers, and the four primer pairs provided by the invention can be directly applied to more lilium plants for germplasm resource and genetic diversity analysis. The four molecular markers disclosed by the invention are all from growth related gene EST sequences of lilies, and a good foundation is established for the cloning of growth related genes of lilium plant flowers and gene sequence analysis.

Description

technical field [0001] The invention relates to the field of molecular biology DNA marker technology and application, in particular to a pair of primers for detection of Lilium EST-SSR. Background technique [0002] Molecular markers are widely used in genetic diversity analysis of germplasm resources and assisted breeding. The research on molecular markers of Lilium began in 1993, but it was limited to the level of scientific and technological development. The markers developed on Lilium plants in the following ten years were mainly random amplified polymorphic DNA (RAPD). Generation of molecular markers, which have poor reproducibility. The expressed sequence tag (Express sequence tag, EST) contains many simple sequence repeats (Simple sequence repeat, SSR), which has become a generally recognized second-generation molecular marker, with the characteristics of good repeatability, co-dominance, and easy detection. [0003] In 2008 and 2011, Yang Suli and Sung-Il Lee deve...

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 Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 杜方高中山夏宜平
Owner ZHEJIANG UNIV