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Excited Information and Courses from MediaLab, Inc.

These are the MediaLab courses that cover Excited and links to relevant pages within the course.

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Fundamentals of Molecular Diagnostics
Terms and Definitions

Term Definition Codon A three nucleotide base sequence that codes for an amino acid Genome The genetic code composed of 64 codons that code for 21 amino acids and 3 stop codons. (amino acids are the building blocks of proteins and stop codons stop the writing process much like a period at the end of a sentence) Nucleic acid Polymer made of monomers; two examples are RNA and DNA Transcription Process of transferring information from DNA into an RNA message Translation The formation of an amino acid from RNA Deoxyribonucleic Acid (DNA) A double-stranded polymer of nucleotides that houses genetic information Ribonucleic acid (RNA) Typically a single-stranded polymer that is much shorter than DNA but chemically similar with a few differences (e.g.- uracil replaces thymine). Replication Reproduction of DNA content from parent to daughter cell during cell division Amplification methods Techniques that increase the amount of the target, the detection signal, or the probe so that sequences are readily detected Fluorescence The emission of light at a longer wavelength when the light is excited at a shorter wavelength Oligonucleotide Short single-stranded nucleic acid Probe A nucleic acid used to identify a hybridization target Polymerase Chain Reaction (PCR) An amplification method performed in vitro

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Detection

Detection techniques can vary in both direct and amplified methodologies and can include labeling either the probe or the target molecule of interest:Chemiluminescence: Release of light energy at the end of a chemical reaction that is detected by a luminometer. Uses a label such as acridinium ester. Electrophoresis: movement in a matrix such as a gel that is caused by an electrical field.Enzyme: Uses enzyme and substrate principles to label the appropriate target or probe. Can be combined with fluorescence or dyes for detection.Fluorescence: Molecules that emit light at a longer wavelength when excited at a shorter wavelength. Detection techniques include fluorescent staining of nucleic acids as well as fluorescent labeled probes that are measured in a fluorometer or with fluorescent polarization.Radioactivity: Uses a labeling technique where the radioactive label is then measured in a scintillation counter. The earliest assays utilized radioactive decay.

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Match the following detection techniques with the most appropriate description:View Page

Pharmacology in the Clinical Lab: Therapeutic Drug Monitoring and Pharmacogenomics
FPIA

Fluoresence polarization immunoassay (FPIA) is also a homogenous competitive immunoassay. In this system, fluorescein-labeled drug competes with unlabeled drug from the patient's serum sample for binding sites on an antibody reagent. The patient's sample, presumably containing the therapeutic drug that is being monitored, and the fluorescein-labeled drug are added to a chamber containing antibody for that drug. The labeled and unlabeled drug will compete for binding sites on the antibody. The greater the amount of drug in the sample, the fewer the number of binding sites that are available for the labeled analyte, leaving a greater number of small, free fluorescein-labeled molecules in the solution.When the chamber is excited with plane polarized light, fluorescein will absorb the light and emit it at a higher wavelength as fluorescent light. A small, free fluorescein-labeled drug rotates randomly and faster than it would if it were bound to antibody, interrupting the light and leading to less emission of light. The larger antibody-drug-fluorescein complexes rotate slower and emit more light in the measured plane. A lower level of drug in the patient's sample results in greater emission of polarized light because there are more antibody-drug-fluorescein complexes present to produce light in the measured plane. A higher level of drug in the patient's sample results in a lower emission of polarized light. This inverse relationship between the concentration of the drug and the polarization units (signal) is illustrated in the image below.

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