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

A kind of Acer palmatum internal reference gene and its application

An internal reference gene, Acer palmatum technology, applied in the field of molecular biology

Active Publication Date: 2020-10-27
CROP RES INST OF FUJIAN ACAD OF AGRI SCI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there has been no report on the isolation and identification of the internal reference gene of Acer palmatum Actin. In the study of the quantitative expression of Acer palmatum-related genes using real-time fluorescent quantitative PCR, the stable The research on the expressed internal reference gene is also blank

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
  • A kind of Acer palmatum internal reference gene and its application
  • A kind of Acer palmatum internal reference gene and its application
  • A kind of Acer palmatum internal reference gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The acquisition of internal reference gene of embodiment 1

[0036] Extract RNA first, specifically: use the general plant total RNA extraction kit to extract total RNA, specifically: weigh 0.2g of 'Golden Maple' leaves at the coloring stage in a mortar, add liquid nitrogen and quickly grind to powder Transfer the powder to a 1.5mL RNase free centrifuge tube, add 1mL lysate PL to mix well, incubate at 65°C for 5min, then centrifuge at 4°C and 12000rpm for 10min; take the supernatant and transfer to RNase free filter In the column, centrifuge at 4°C and 10,000 rpm for 1 min, collect the filtrate in a collection tube, add 1 volume of 70% ethanol, and mix well; transfer the mixed solution and the precipitate into the adsorption column RA together, Centrifuge at 4°C and 12000rmp for 1min, discard the waste liquid, put the adsorption column back into the collection tube; add 500uL deproteinized solution RE, centrifuge at 4°C and 12000rmp for 1min, discard the waste liquid; a...

Embodiment 2

[0043] Embodiment 2 real-time fluorescent quantitative PCR design and conventional PCR detection

[0044] Based on the nucleotide sequence of the internal reference gene obtained in Example 1, and following the principles of real-time fluorescent quantitative PCR primer design, a pair of fluorescent quantitative specific primers were designed, the amplified fragment was 162bp, and the pair of fluorescent quantitative specific primers was The real-time fluorescent quantitative PCR primers (as shown in SEQ ID NO: 10, 11): Actin-RT-F: 5'-GGATGCCTATGTTGGTGACG-3', Actin-RT-R: 5'-AGCACTGGGTGTTCCTCTGG-3'.

[0045] The general plant total RNA extraction kit was used to extract the total RNA from the leaves of 'Golden Maple' during the coloring stage, and the first strand of cDNA was synthesized using the method of PrimeScriptTM II 1st Strand cDNA Synthesis Kit (purchased from TaKaRa Company). It is transcribed into cDNA; then the first strand of the obtained cDNA is used as a template...

Embodiment 3

[0049] Embodiment 3 real-time fluorescent quantitative PCR primer verification

[0050] The general plant total RNA extraction kit was used to extract the total RNA from the leaves of 'Golden Maple' during the coloring stage, and the first strand of cDNA was synthesized using the method of PrimeScriptTM II 1st Strand cDNA Synthesis Kit (purchased from TaKaRa Company). Transcribed into cDNA; then use the first strand of the obtained cDNA as a template, and perform real-time fluorescence quantitative analysis according to the method of TaKaRa's SYBR@Premix Ex TaqTM II instructions, and the reaction system and reaction procedure are as follows:

[0051] Reaction system: The total volume of the reaction system is 10 μL, including 5.0 μL of 2×SYBRⅡ Premix Ex Taq TM , the upstream primer (10 μ mol / L) of fluorescent quantitative PCR primer in 0.4 μ L embodiment 2, the downstream primer (10 μ mol / L) of fluorescent quantitative PCR primer in 0.4 μ L embodiment 2, 0.2 μ L ROX Reference ...

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 provides a Japanese maple reference gene, and also discloses a real-time fluorescence quantification PCR primer by using the Japanese maple reference gene as a basic design. The currentsituation that existing Japanese maple quantification PCR detection is free of a reference gene is solved; moreover, when the designed real-time fluorescence quantification PCR primer is used for analysis of Japanese maple reference gene expression, the stability, reliability and repeatability of analysis and research of Japanese maple gene expression can be improved; and moreover, the designed real-time fluorescence quantification PCR primer is applicable for related gene quantification expression research of different leaf stages and different leaf color varieties of Japanese maple, so thatthe accuracy of obtained data can be obviously improved, and technical support is provided for reducing the error of different samples, standardizing total RNA quantity and guaranteeing the reliability of data.

Description

【Technical field】 [0001] The invention belongs to the technical field of molecular biology, and specifically relates to an acer palmatum gene and its application. Specifically, a pair of fluorescent quantitative specific primers are designed based on the nucleotide sequence of the acer palmatum gene. 【Background technique】 [0002] Acer palmatum (Acer palmatum) is a deciduous small tree belonging to the family Aceraceae (Aceraceae) and is a foliage tree species with high ornamental value. There are many varieties of Acer palmatum, including 'Golden Maple' (golden leaves), 'Orange Dream' (orange leaves with light red edges), 'Gui' (yellow green leaves), 'Butterfly Dance' (green leaf edges Pink), 'Qingzhiwei' (green leaves), 'Green Duck' (green leaves), 'Bugatti' (purple leaves), 'Sanjihong' (deep purple leaves), 'Kasayama' ( Brick red leaves, red veins), 'Goddess' (purple red leaves, red veins), 'Out of Orangutan' (dark red leaves), etc., the leaf color changes in a variety ...

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): C12N15/29C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/158C12Q2600/166
Inventor 林榕燕钟淮钦陈裕德林兵罗远华
Owner CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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