Quantifying Radioactive Transcripts
Quantifying Radioactive Transcripts - Related Videos
03:18
Radioactivity and Nuclear Equations
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2020
Nuclear chemistry is the study of reactions that involve changes in nuclear structure. The nucleus of an atom is composed of protons and, except for hydrogen, neutrons. The number of protons in the nucleus is called the atomic number (Z) of the element, and the sum of the number of protons and the number of neutrons is the mass number (A). Atoms with the same atomic number but different mass numbers are isotopes of the same element. A nuclide of an element has a specific number of protons and...
02:48
Radioactive Decay and Radiometric Dating
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2020
Radioactivity is a spontaneous disintegration of an unstable nuclide and is a random process, as all the nuclei in the sample do not decay simultaneously. The number of disintegrations per unit time is called the activity (A), which is directly proportional to the number of nuclei in the sample. The decay constant (λ) is an average probability of decay per nucleus in unit time. The SI unit for activity is the becquerel, which is one disintegration per second. Another unit of activity is the...
11:19
Merging Absolute and Relative Quantitative PCR Data to Quantify STAT3 Splice Variant Transcripts
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Cited by 2 •
2016
Tandem splicing events occur at sites less than 12 nucleotides apart. Quantifying ratios of such splice variants is feasible using an absolute quantitative PCR approach. This manuscript describes how splice variants of the gene STAT3, in which two splicing events results in Serine-701 inclusion/exclusion and α/β C-termini, can be quantified.
04:25
Substrate Oxidation Assay in Cultured Cells: An In Vitro Assay to Quantify Substrate Oxidation in Cells by Measuring Radioactive Signals from Trapped CO2
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2025
This video demonstrates a quantification technique for CO2 released during substrate oxidation using 14C-radiolabeled substrates. The released 14CO2 is trapped in an alkaline solution and quantified by a scintillation counter. The oxidation of different substrates varies between tissues and reflects the pathophysiological condition of the tissue.
03:23
Types of Radioactivity
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2020
The most common types of radioactivity are α decay, β decay, γ decay, neutron emission, and electron capture. Alpha (α) decay is the emission of an α particle from the nucleus. For example, polonium-210 undergoes α decay: Alpha decay occurs primarily in heavy nuclei (A > 200, Z > 83). Loss of an α particle gives a daughter nuclide with a mass number four units smaller and an atomic number two units smaller than those of the parent nuclide. Beta (β) decay is the emission of an electron or...
FAQs
The measured signal depends on how efficiently the radioactive nucleotide precursor enters newly synthesized RNA and how consistently the resulting radioactivity is detected. When labeling and detection conditions remain controlled, signal intensity provides a basis for comparing transcript abundance or transcriptional activity between samples. Changes in signal therefore require interpretation alongside the experimental controls.
Controlled conditions make differences in radioactivity more likely to reflect biological changes rather than variation in labeling or measurement. Consistency is especially important when comparing transcription between experimental conditions, because signal intensity is used as an indicator of transcript abundance. Poorly controlled conditions can make apparent differences difficult to attribute to gene regulation.
These techniques provide different ways to detect or measure the radioactive signal associated with RNA. Gel analysis can support examination of separated transcripts, hybridization can help isolate or detect particular transcript populations, and scintillation counting measures radioactivity quantitatively. The appropriate readout depends on whether the experiment emphasizes transcript separation, identification, or overall radioactive signal.
A typical workflow introduces a radioactive nucleotide precursor during RNA synthesis, then separates or isolates the resulting transcripts before measuring their radioactivity. Detection may use gel analysis, hybridization, or scintillation counting. Researchers compare the resulting signal under controlled labeling and detection conditions to evaluate relative transcript abundance or transcriptional activity.
This approach can examine promoter activity, transcription kinetics, RNA processing, and transcript stability. Promoter studies address regulation of RNA production, whereas kinetic analyses focus on changes in transcriptional activity over an experiment. Measurements related to processing or stability extend the analysis beyond production alone, helping distinguish how gene expression is regulated after transcription begins.
Quantifying radioactive transcripts is useful when experiments need a sensitive comparison of RNA production across different biological conditions. Differences in measured signal can indicate altered transcriptional activity when labeling and detection remain controlled. In biology, the method therefore supports assessment of regulatory effects at the transcript level and helps connect experimental conditions with changes in gene expression.