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Mixtures of Acids03:27

Mixtures of Acids

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The pH of a solution containing an acid can be determined using its acid dissociation constant and its initial concentration. If a solution contains two different acids, then its pH can be determined using one of several methods depending upon the relative strength of the acids and their dissociation constants.
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The pH of a solution containing an acid can be determined using its acid dissociation constant and initial concentration. If a solution contains two different acids, then its pH can be determined using one of several methods depending on the relative strength of the acids and their dissociation constants.
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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
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Full-Length Multi-Barcoding: DNA Barcoding from Single Ingredient to Complex Mixtures.

Peng Zhang1,2, Chunsheng Liu3, Xiasheng Zheng4

  • 1School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China. zheng.x.s1987@163.com.

Genes
|May 10, 2019
PubMed
Summary

Full-length multi-barcoding (FLMB) using long-read sequencing accurately identifies species in complex Traditional Chinese Medicine formulas. This novel DNA barcoding method overcomes limitations of previous technologies for authenticating multi-species mixtures.

Keywords:
DNA barcodingITS2Sheng-Mai-San (SMS)multi-species mixturesnext-generation sequencingsingle-molecule real-time (SMRT)

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Area of Science:

  • Molecular Biology
  • Genomics
  • Pharmacognosy

Background:

  • Traditional Chinese Medicine (TCM) often involves multi-species herbal formulas, making accurate identification critical for clinical use.
  • Existing DNA barcoding methods, particularly Next-Generation Sequencing (NGS), face challenges with short reads and systematic errors, limiting their application in complex mixtures.
  • Previous DNA barcoding applications have predominantly focused on single-species identification.

Purpose of the Study:

  • To develop and validate a novel Full-length multi-barcoding (FLMB) method utilizing long-read sequencing for authenticating biological compositions in herbal compound formulas.
  • To assess the sensitivity, accuracy, and reliability of the FLMB method in identifying species within complex mixtures.
  • To analyze the composition of classical TCM formulas Sheng-Mai-San (SMS) and Sanwei-Jili-San (SJS) using the FLMB approach.

Main Methods:

  • Employed Full-length multi-barcoding (FLMB) with single-molecule real-time (SMRT) long-read sequencing technology.
  • Sequenced full-length amplicons of ITS2 and *psbA-trnH* for SMS, and ITS2 and CO1 for SJS.
  • Utilized clone-dependent Sanger sequencing as a parallel control method for comparison.

Main Results:

  • Successfully authenticated all ingredients in the Sheng-Mai-San (SMS) and Sanwei-Jili-San (SJS) samples at the species level.
  • Identified 11 exogenous species, including common contaminants, within the analyzed herbal formulas.
  • Methodology analysis confirmed FLMB as a sensitive, accurate, and reliable approach for complex mixture identification.

Conclusions:

  • Full-length multi-barcoding (FLMB) via long-read sequencing is a superior and feasible method for identifying biological components in complex mixtures.
  • The established FLMB approach provides a robust solution for DNA barcoding applications in multi-species herbal formulas.
  • This technique enhances the authentication of Traditional Chinese Medicine, ensuring clinical safety and efficacy.