Pure honeysuckle polymerase chain reaction-restriction map identification method

A technology of restriction endonuclease and honeysuckle, which is applied in the field of quality identification of Chinese medicinal material germplasm, can solve the problems such as inability to distinguish honeysuckle in shape and microscopic pattern, achieve a reliable identification method and ensure the quality of medicinal materials.

Inactive Publication Date: 2003-08-27
CHINA PHARM UNIV
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the honeysuckle in the non-genuine area and the authentic area

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
  • Pure honeysuckle polymerase chain reaction-restriction map identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0014] a. DNA extraction: proceed as usual, and adjust the DNA concentration of the test sample to 0.1-0.3 μg / μl with deionized water;

[0015] a. Polymerase chain reaction:

[0016] (1) For polymerase chain reaction, 30 μl is used as a reference, and the dosages of various items are as follows:

[0017] 10×PCR buffer 3μl

[0018] MgCl 2 (25mM) 2.4μl

[0019] dNTP (each 10mM) 0.6μl

[0020] Primers LjP1, LjP2 (30pmol / μl) 0.5μl each

[0021] 1~2 μl DNA template for test product

[0022] TaqDNA polymerase 1U (0.2μl)

[0023] Deionized water make up to 30μl

[0024] After mixing, centrifuge at high speed for a few seconds

[0025] (2) PCR reaction conditions: the reaction is carried out on a PCR instrument, and the reaction conditions are:

[0026] Pre-denature at 97°C for 4 minutes, and then carry out 30 cycles according to the following conditions:

[0027] Denaturation at 97°C for 15 seconds

[0028] Refolding at 53°C for ...

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 polymerase chain reactino-restrictive enzyme-cut chart molecule (PCR-RFLP) method accurately identifying honeysuckle in genuine producing areas (Henan, Shandong) and nongenuine producing areas (other producing areas). The characteristic DNA alkali sequence of honeysuckle in genuine: 5'-GCCCCCCCGC CCCGCCTCCC ACAGGGTCGC G-3'; restrictive inner cut enayme EcoNi and its same cut-enzyme can identify and cut this sequence. The identifying course: extracting DNA of medical materials; using a pair of guide matters to make PCR increase; using restrictive inner cut enayme EcoNI and its same cut enayme to digest PCR product; analyzing the result by using gelation and electrophoresis of gelose; in contrast with the characteristic result of PCR-RFLP of genuine honeysuckle, judging if the tested product is genuine honeysuckle.

Description

1. Technical field: [0001] The invention relates to the technical field of quality identification of germplasm of traditional Chinese medicinal materials. 2. Background technology: [0002] Honeysuckle is the dry flower bud or the first flower of Lonicera japonica Thunb., a plant of Lonicera japonica Thunb. It has strong antibacterial effect and has good curative effect on various acute and chronic inflammations. The medicinal materials of honeysuckle are obviously authentic. For example, through our preliminary determination of the content of chlorogenic acid, the content of chlorogenic acid in Henan and Shandong honeysuckle in authentic areas is generally higher than that in other non-authentic areas, which shows that the quality of medicinal materials is related to the region, habitat, etc. closely related. However, honeysuckle from non-genuine areas and authentic areas cannot be distinguished by morphological and microscopic characteristics. Due to the isolation of geo...

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): C07H21/04C12Q1/24C12Q1/68
Inventor 李萍王冲之周开亚
Owner CHINA PHARM UNIV
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