Quality control method for detecting agkistrodon component in Jinlong capsule

A quality control method, the technology of Jinlong Capsule, is applied in the field of molecular biology, which can solve the problem of inability to detect fluorescent signals, and achieve the effect of high application value, high accuracy and good repeatability.

Inactive Publication Date: 2018-06-05
SOUTHERN MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the probe is intact, the fluorescent signal emitted by the reporter group is absorbed by the quencher group, and the fluorescent signal cannot be detected

Method used

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  • Quality control method for detecting agkistrodon component in Jinlong capsule
  • Quality control method for detecting agkistrodon component in Jinlong capsule
  • Quality control method for detecting agkistrodon component in Jinlong capsule

Examples

Experimental program
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Effect test

Embodiment 1

[0035] 1. DNA extraction system

[0036] The DNA extraction of the sample was carried out using the improved SDS extraction method, and the required reagents included: buffer A (1M Tris-HCL; 5M NaCL; 0.5M EDTA; sodium dodecyl sulfate), buffer B (3mol / L sodium acetate ), buffer C (2mg / mL proteinase K). The steps are as follows: Take the contents of 2 Jinlong capsules, add 2ml of buffer A, 100μL of buffer C, shake and mix well, bathe in water at 65°C for 1.5h, add 500μL of buffer B, 2mL of chloroform-isoamyl alcohol (24:1) Extract, add an equal volume of isoamyl alcohol to the supernatant to precipitate at -20°C for 3 hours, centrifuge at 12,000 rpm for 20 minutes, wash the precipitate with 75% ethanol, dry in the air, add TE to dissolve, purify DNA with Tiangen Purification and Recovery Kit, and follow the PCR product Purification steps are carried out.

[0037] 2. System and procedures

[0038] For the sample to be tested, 20 μl of the reaction system mixed with the probe pri...

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Abstract

The invention provides a quality control method for detecting an agkistrodon component in a Jinlong capsule. The quality control method comprises the following steps: extracting DNA in a Jinlong capsule sample; designing and synthesizing a Taqman probe; establishing a Taqman qPCR standard curve; and carrying out Taqman qPCR detection. According to the quality control method, the agkistrodon component in the Jinlong capsule can be accurately, reliably and effectively authenticated.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a quality control method for detecting Aggresis components in Jinlong Capsules by qPCR based on Taqman probes. Background technique [0002] Jinlong Capsule is the first anti-cancer fresh animal drug approved by the state. Its prescription is composed of fresh Shougong, fresh white snake and fresh Viper in a ratio of 2:1:1. , can inhibit tumor growth, inhibit postoperative local recurrence and distant metastasis, regulate the body's immunity, and be used for the treatment of primary liver cancer, lung cancer and other cancers and adjuvant chemotherapy and radiotherapy. Among them, Viper is a precious Chinese medicinal material, which has the effects of dispelling wind, dredging collaterals, and relieving spasm. In recent years, the medicinal value and economic value of snakes have increased significantly, and the social demand continues to rise. With the gradual sca...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6806C12Q1/686C12N15/10
CPCC12N15/1003C12Q1/6806C12Q1/686C12Q1/6888C12Q2561/113C12Q2563/107C12Q2545/114C12Q2561/101
Inventor 晁志叶浩婷田恩伟
Owner SOUTHERN MEDICAL UNIVERSITY
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