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Related Concept Videos

Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

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RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
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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

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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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Free-Radical Chain Reaction and Polymerization of Alkenes02:35

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The conversion of alkenes to macromolecules called polymers is a reaction of high commercial importance. The structure of the polymer is defined by a repeating unit, while the terminal groups are considered insignificant. The average degree of polymerization represents the number of repeating units in the polymer molecule and is denoted by the subscript n.
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Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
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Electron Transport Chains01:28

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The final stage of cellular respiration is oxidative phosphorylation that consists of two steps: the electron transport chain and chemiosmosis. The electron transport chain is a set of proteins found in the inner mitochondrial membrane in eukaryotic cells. Its primary function is to establish a proton gradient that can be used during chemiosmosis to produce ATP and generate electron carriers, such as NAD+ and FAD, that are used in glycolysis and the citric acid cycle.
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Gastric Mucosa Quantitative Polymerase Chain Reaction Analysis for Detecting Helicobacter pylori and Antibiotic Resistance
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Gastric Mucosa Quantitative Polymerase Chain Reaction Analysis for Detecting Helicobacter pylori and Antibiotic Resistance

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Quantitative polymerase chain reaction.

Stuart N Peirson1, Jason N Butler

  • 1Division of Circadian and Visual Neuroscience, Univesity of Oxford, UK.

Methods in Molecular Biology (Clifton, N.J.)
|April 10, 2007
PubMed
Summary
This summary is machine-generated.

Quantitative PCR (qPCR), a real-time PCR method, enables high-throughput gene expression analysis. This guide covers essential principles for reliable qPCR assay setup and troubleshooting.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Real-time quantitative PCR (qPCR) is widely adopted due to advancements in real-time PCR platforms.
  • qPCR allows monitoring of amplification products using fluorescent dyes, enabling measurements during the exponential phase.
  • The technique offers high-throughput analysis of multiple transcripts from small samples with high dynamic range and sensitivity.

Purpose of the Study:

  • To provide fundamental principles for establishing a quantitative real-time PCR assay.
  • To address common challenges and troubleshooting for new users of qPCR technology.
  • To focus on universal principles rather than specific platforms or chemistries.

Main Methods:

  • Discussion of core principles including sample preparation and experimental design.
  • Emphasis on the use of internal controls and assay considerations.
  • Exploration of data analysis approaches for qPCR experiments.

Main Results:

  • qPCR facilitates efficient analysis of gene expression from limited biological material.
  • Reliable data acquisition may require optimization and troubleshooting due to technical variations.
  • The article aims to demystify the process for new researchers.

Conclusions:

  • Understanding basic principles is crucial for successful qPCR assay implementation.
  • This guide serves as a foundational resource for setting up and troubleshooting qPCR experiments.
  • The technique's power lies in its sensitivity and throughput when applied correctly.