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Related Concept Videos

Phase I Reactions: Hydrolytic Reactions01:15

Phase I Reactions: Hydrolytic Reactions

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Hydrolysis, a cornerstone of phase I biotransformation reactions, uses water to cleave chemical bonds. This process is pivotal in drug metabolism, generating more polar metabolites that can be easily excreted.
An important hydrolytic reaction is ester hydrolysis. Ester bonds, often found in prodrugs, are broken down, increasing the solubility of drugs like aspirin and lidocaine for more straightforward elimination. Amide hydrolysis is another critical reaction, targeting amide bonds prevalent...
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Phase II Reactions: Acetylation Reactions01:24

Phase II Reactions: Acetylation Reactions

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Acetylation, a phase II biotransformation reaction, introduces an acetyl group to drugs or their metabolites. Acetyltransferase enzymes facilitate this reaction, which resembles α-amino acid conjugation due to the addition of a functional group to the drug molecule.
The substrates for acetylation are typically drugs or their metabolites with an amino, sulfonamide, or hydrazine functional group. Acetylation can occur at several points in the drug molecule, including primary, secondary, and...
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Phase I Reactions: Reductive Reactions01:27

Phase I Reactions: Reductive Reactions

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Phase I biotransformation reductive reactions are chemical processes that modify drugs by introducing or revealing polar functional groups via reduction. Enzymes called reductases catalyze these reactions, playing a pivotal role in drug metabolism by transforming lipophilic drugs into more polar, water-soluble metabolites for easy excretion. An essential type of reductive reaction is the carbonyl group reduction, where aldehydes and ketones are reduced to alcohols. An example is the...
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Phase II Reactions: Methylation Reactions01:17

Phase II Reactions: Methylation Reactions

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Methylation is a phase II biotransformation process involving the attachment of a methyl group to a substrate. Enzymes known as methyltransferases orchestrate this reaction.
The mechanism of methylation unfolds in two stages. The first stage sees a methyltransferase enzyme facilitating the transfer of a methyl group from S-adenosylmethionine (SAM) to the substrate, forming S-adenosylhomocysteine (SAH). The second stage involves further metabolism of SAH into homocysteine, which can be recycled...
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

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Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
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Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies
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Microdevice-based solid-phase polymerase chain reaction for rapid detection of pathogenic microorganisms.

Quang Nghia Pham1, Kieu The Loan Trinh1, Seung Won Jung2

  • 1Department of BioNano Technology, Gachon University, Seongnam-si, Gyeonggi-do, Republic of Korea.

Biotechnology and Bioengineering
|May 20, 2018
PubMed
Summary

This study presents a lab-on-a-chip microdevice for rapid DNA analysis. The solid-phase polymerase chain reaction (SP-PCR) platform enables quick identification of foodborne pathogens and harmful algae at the point-of-need.

Keywords:
algafoodborne pathogenintegrated microdeviceon-chip detectionpoint-of-need (PON) testingsolid-phase polymerase chain reaction (SP-PCR)

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Molecular Diagnostics

Background:

  • Point-of-need (PON) DNA analyses require integrated amplification and detection.
  • Lab-on-a-chip devices offer potential for portable molecular diagnostics.

Purpose of the Study:

  • To develop and demonstrate a lab-on-a-chip microdevice for solid-phase polymerase chain reaction (SP-PCR).
  • To enable rapid, integrated DNA amplification and detection for point-of-need applications.

Main Methods:

  • Fabrication of a polycarbonate microdevice with microchambers via thermal bonding.
  • Modification of microchambers with polyethyleneimine and glutaraldehyde for primer immobilization.
  • Implementation of SP-PCR using immobilized forward primers and free reverse primers for amplicon generation and detection.

Main Results:

  • Successful generation of target amplicons covalently attached to microchambers.
  • Direct identification of foodborne pathogens (Salmonella spp., Staphylococcus aureus) and harmful alga (Cochlodinium polykrikoides).

Conclusions:

  • The developed SP-PCR microdevice provides a versatile platform for rapid, integrated DNA analysis.
  • This technology is suitable for point-of-need testing of foodborne pathogens and environmental biogenic targets.