Method and PCR (polymerase chain reaction) reagent kit for identifying DNA (deoxyribonucleic acid) barcodes of animal medicinal materials

A technology of animal medicinal materials and kits, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of low genomic DNA abundance and high genomic heterogeneity, and achieve the effect of wide application and effective amplification

Inactive Publication Date: 2015-12-02
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the shortcomings of high genome heterogeneity, low genomic DNA abundance, and PCR reaction inhibitors among Chinese medicinal material samples, the present invention provides a method for identifying animal medicinal materia...

Method used

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  • Method and PCR (polymerase chain reaction) reagent kit for identifying DNA (deoxyribonucleic acid) barcodes of animal medicinal materials
  • Method and PCR (polymerase chain reaction) reagent kit for identifying DNA (deoxyribonucleic acid) barcodes of animal medicinal materials
  • Method and PCR (polymerase chain reaction) reagent kit for identifying DNA (deoxyribonucleic acid) barcodes of animal medicinal materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] PCR reaction system:

[0032]

[0033] Among them, the primers are:

[0034] Primer R: GGTCAACAAATCATAAAGATATTGG

[0035] Primer F: TAAACTTTTCAGGGTGACCAAAAAAATCA

[0036] Wherein, the enhancer includes: bovine serum albumin 80 micrograms per microliter; dithiothreitol 10 micromoles per microliter; 0.5% NP40; betaine 1 mole per microliter.

[0037] PCR reaction program setting

[0038]

[0039] Goto: 2for5cycles

[0040] 94℃30s

[0041] 50℃30s

[0042] 72℃1min

[0043] Goto: 6for35cycles

[0044] 72℃5min

[0045] END

Embodiment 2

[0052] PCR reaction system:

[0053]

[0054] Among them, the primers are:

[0055] Primer R: GGTCAACAAATCATAAAGATATTGG

[0056] Primer F: TAAACTTTTCAGGGTGACCAAAAAAATCA

[0057] Wherein, the enhancer includes: bovine serum albumin 85 micrograms per microliter; dithiothreitol 8 micromoles per microliter; 0.5% NP40; betaine 0.8 moles per microliter.

[0058] PCR reaction program setting

[0059]

[0060]

[0061] Goto: 2for5cycles

[0062] 94℃30s

[0063] 50℃30s

[0064] 72℃1min

[0065] Goto: 6for35cycles

[0066] 72℃5min

[0067] END

Embodiment 3

[0073] PCR reaction system:

[0074]

[0075]

[0076] Among them, the primers are:

[0077] Primer R: GGTCAACAAATCATAAAGATATTGG

[0078] Primer F: TAAACTTTTCAGGGTGACCAAAAAAATCA

[0079] Wherein, the enhancer includes: bovine serum albumin 75 micrograms per microliter; dithiothreitol 10 micromoles per microliter; 0.4% NP40; betaine 1.1 moles per microliter.

[0080] PCR reaction program setting

[0081]

[0082] Goto: 2for5cycles

[0083] 94℃30s

[0084] 50℃30s

[0085] 72℃1min

[0086] Goto: 6for35cycles

[0087] 72℃5min

[0088] END

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Abstract

The invention provides a method and a PCR (polymerase chain reaction) reagent kit for identifying DNA (deoxyribonucleic acid) barcodes of animal medicinal materials. The PCR reagent kit comprises PCR enhancers. The PCR enhancers comprise bovine serum albumin, dithiothreitol, betaine and nonidet p40. The method and the PCR reagent kit for identifying the DNA barcodes of the animal medicinal materials have the advantages that low-abundance traditional Chinese medicine samples with difficulty in amplification, particularly, samples with difficulty in identifying DNA barcodes, such as samples of buffalo horns, antelope horns, tortoise shells, carapax trionycis, pangolins and snake slough, can be effectively amplified, accordingly, the amplification success rate which is close to 100%, of DNA barcode identification techniques can be guaranteed, and DNA barcode techniques can be widely applied.

Description

technical field [0001] The invention relates to a PCR kit, in particular to a PCR kit and an identification method which can be used for identification of DNA barcodes of pharmacopoeia animal medicinal materials. Background technique [0002] The identification of traditional Chinese medicine is the premise and basis for studying the variety and quality of traditional Chinese medicine, formulating the standard of traditional Chinese medicine, and finding and expanding the source of medicine. The four traditional identification methods of traditional Chinese medicine are genetic identification, trait identification, microscopic identification and physical and chemical identification. The theoretical basis of these traditional identification methods is based on the analysis of the traits of taxa. Results are susceptible to subjective and objective factors. [0003] Genomic DNA sequence is the basis and determinant of the morphology and basic properties of species, as well as ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q1/6895
Inventor 石林春宋经元庞晓慧姚辉韩建萍陈士林
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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