A primer and method for identifying yellow-flowered ox ears and ox ears using EST microsatellite markers

A technology of yellow ox ears and microsatellite markers, which can be used in biochemical equipment and methods, determination/inspection of microorganisms, DNA/RNA fragments, etc., and can solve the problems of few applications and low medicinal value.

Active Publication Date: 2022-02-01
GUILIN NORMAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The flower color and flower shape of the yellow-flowered cow's ear are very beautiful, and it can be developed into an excellent ornamental potted flower. However, compared with the cow's ear, its medicinal value is not high, and it is less used as a Chinese herbal medicine among the people.
However, there is no relevant report on the primers and methods of using EST microsatellite markers to identify yellow-flowered ox ears and ox ears

Method used

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  • A primer and method for identifying yellow-flowered ox ears and ox ears using EST microsatellite markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Constructing and sequencing the transcriptome library of yellow-flowered ox ears and ox ears

[0027] Firstly, the total RNA of yellow-flowered cow's ear and cow's ear is extracted, and the extracted RNA is subjected to agarose gel electrophoresis, nucleic acid concentration and quality detection, and the total RNA concentration is judged. The mRNA containing poly(A) was enriched and purified by magnetic beads containing Oligo(dT), double-stranded cDNA was reverse transcribed and purified, poly(A) was added using Klenow and sequencing adapters were connected, and agarose gel was used to Gel electrophoresis for the recovery and purification of cDNA fragments of the desired size. The cDNA fragments in the PCR amplification products were used, and the PCR target products were gel-recovered, and the qualified cDNA fragments were purified and detected, and sequenced on Illumina HiseqTM2500 as a cDNA library. The sequencing peak map was read by Base calling softwa...

Embodiment 2

[0032] Embodiment 2: the method for identifying yellow-flowered ox ears and ox ears with the primers determined in embodiment 1

[0033] (1) The extraction of the yellow-flowered cow's ear genome DNA and the cow's ear genome DNA is carried out as follows:

[0034] 1) Preheat the 10mL centrifuge tube with 4mL CTAB extract and 80μl mercaptoethanol in a 65°C water bath;

[0035] 2) Take 1-2g of plant leaves in a mortar, grind an appropriate amount of liquid nitrogen into powder, then transfer to preheated CTAB extract, mix well, keep warm in a water bath at 65°C for 30 minutes, during this period, shake it every 5-6 minutes;

[0036] 3) Cool the warmed sample on ice to room temperature quickly, add an equal volume of chloroform:isoamyl alcohol (24:1), mix thoroughly and centrifuge at 12000rpm for 10min, take 500-1000μL of the supernatant into a 1.5ml centrifuge tube medium (the rest of the supernatant was stored at -20°C for later use);

[0037] 4) Add 100 μL of silica bead sus...

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Abstract

The invention discloses a primer and a method for identifying yellow-flowered ox ears and ox ears by using EST microsatellite markers. The primers are EST-F: 5'-AACGGGTTGTGTTCTGACGT-3'; EST-R: 5'-CGATTCGAGACCCCCTTCG-3'. The method for using EST microsatellite markers to identify yellow-flowered ox ears and ox ears is as follows: first prepare a genomic DNA solution, and use this as a template to perform PCR amplification on the genomic DNA with the aforementioned primers, and perform polyacrylamide gel on the resulting amplification products For electrophoresis, when the electrophoresis pattern shows a band of 230bp, it is a cow's ear; when the electrophoresis pattern shows a band of 210bp, it is a yellow ox ear. The primers and method provided by the invention provide simple and stable molecular markers for identifying the yellow-flowered ox ear and the ox ear, and solve the problem of distinguishing the yellow-flowered ox ear and the ox ear.

Description

technical field [0001] The invention relates to a method for identifying plants by using EST microsatellite markers, in particular to a primer and a method for identifying yellow-flowered ox ears and ox ears by using EST microsatellite markers. Background technique [0002] Primulina lutea (Primulina lutea) and Chirita eburnea Hance (Chirita eburnea Hance) are both plants of the genus Primulus in Gesneriaceae. They are similar in shape and have a very close relationship. In the past classification system, Primulina eburnea Hance was considered only A variety of cow's ear that was not renamed until it was identified in 2004. [0003] In addition to the difference in flower morphology and adaptability, the chemical components contained in the rhizomes and leaves of the two Gesneria plants are also different. Among them, Cow's ear contains a variety of phenylethanol glycosides, but in Cow's ear However, the above-mentioned types of substances were not detected in . In folklor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 李博
Owner GUILIN NORMAL COLLEGE
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