Paphiopedilum DNA extraction buffer, and preparation method and use method thereof

A technology of buffer solution and Paphiopedilum, applied in the field of genetic engineering, can solve problems such as viscosity and difficulty in aspiration of supernatant

Active Publication Date: 2019-04-12
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The leaves of Paphiopedilum apricot are thicker and have a longer lifespan. Therefore, the leaf tissue contains more pigments and polysaccharides. During the extraction process, there will often be viscous gelatinous substances, which will cause great damage to the suction of the supernatant. big difficulty

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
  • Paphiopedilum DNA extraction buffer, and preparation method and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Recipe and preparation of Paphiopedilum DNA extraction buffer:

[0035] Ingredients (content per liter):

[0036] 1. 1M Tris-HCl (pH=8.0) 200mL;

[0037] 2. NaCl 14.61g;

[0038] 3. 0.5M EDTA (pH=8.0), 100mL;

[0039] 4. β-mercaptoethanol, 10mL.

[0040] Preparation: Add the above components in 600mL sterile purified water in turn, dissolve and dilute to 1000mL, and store in a refrigerator at 4°C.

Embodiment 2

[0042] Recipe and preparation of Paphiopedilum DNA extraction buffer:

[0043] Ingredients (content per liter):

[0044] 1. 1M Tris-HCl (pH=8.0) 250mL;

[0045] 2. NaCl 18.5g;

[0046] 3. 0.5M EDTA (pH=8.0), 120mL;

[0047] 4. β-mercaptoethanol, 15mL.

[0048] Preparation: Add the above components in 600mL sterile purified water in turn, dissolve and dilute to 1000mL, and store in a refrigerator at 4°C.

Embodiment 3

[0050] 1. Rapid DNA extraction method of Paphiopedilum apricot

[0051] 1. Take the fresh leaves of P. chinensis, freeze them 3 to 4 times in liquid nitrogen, and grind them rapidly. Take 2 scoops of powder in a 5mL centrifuge tube and put them in ice cubes.

[0052] 2. Add 2 mL of the Paphiopedilum DNA extraction buffer prepared in Example 1, mix well, and ice-bath for 10 min.

[0053] 3. Centrifuge (12000rpm) for 5-10min, and discard the supernatant.

[0054]5. Take out the tube, let it cool down to room temperature naturally, add 2 mL of 24:1 (chloroform:isoamyl alcohol = 24:1) to mix, and shake gently continuously for 5-10 minutes.

[0055] 6. Centrifuge (12000rpm) for 10min, absorb 1.8mL of supernatant, add 2mL of 24:1 to mix, shake gently continuously for 5-10min. Centrifuge (12000rpm) for 10min, and absorb 1.6mL of supernatant.

[0056] 7. Add 1.1 mL of 70% (volume ratio) isopropanol, mix well, and place in a -20°C refrigerator for 30 minutes (settling DNA).

[0057...

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 present invention provides a Paphiopedilum DNA extraction buffer, and a preparation method and use method thereof, and belongs to the technical field of genetic engineering. The Paphiopedilum DNAextraction buffer uses water as a solvent, and each liter of the buffer includes: 150-250 mL of Tris-HCl solution, 10-20 g of sodium chloride, 80-120 mL of EDTA solution and 5-15 mL of beta-mercaptoethanol. The Paphiopedilum DNA extraction buffer provided by the invention can remove pigments and polysaccharide substances in a sample, better-quality DNA is obtained and further a main band is obvious when a gel is imaged, while DNA extracted without use of the Paphiopedilum DNA extraction buffer does not show an obvious main band during gel imaging.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and in particular to a paphiopedilum DNA extraction buffer, a preparation method and a use method thereof. Background technique [0002] Paphiopedilum plants have unique flower shapes, rich flower colors and long-lasting flowering period. They are important ornamental flowers in the world and have huge market potential. Due to over-collection and habitat destruction, many species are hard to find in the wild, and all species of Paphiopedilum are included in the protection category of Appendix 1 of the Convention on International Trade in Endangered Species of Wild Fauna and Flora. Paphiopedilum armeniacum is the flagship species of the genus Paphiopedilum, with large, round, golden-yellow flowers. Once discovered, it amazed the entire orchid world and won the first prize at the Tokyo International Orchid Show. Because of its high ornamental value and strong market demand, illegal col...

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
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 黄家林张石宝胡虹
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap