Technology
Multiplex PCR Guide: Panel Design, Use Cases & Performance Trade-offs
Learn how multiplex PCR balances panel breadth, sensitivity, cross-reactivity and instrument compatibility across clinical applications.
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Technology
Learn how multiplex PCR balances panel breadth, sensitivity, cross-reactivity and instrument compatibility across clinical applications.
Technology
A type of high-throughput, multiplex PCR method used to determine abnormal DNA copy number changes of up to 50 genomic DNA or RNA sequences with a single pair of PCR primers. It is frequently used in the detection of genetic aberrations in tumors, and can distinguish sequences differing in only a single nucleotide.
Technology
A technique designed to rapidly enrich and amplify targets derived from samples that yield low levels of nucleic acids, such as formalin-fixed paraffin-embedded (FFPE) tissues, for use in next-generation sequencing. Briefly, total nucleic acid or genomic DNA (gDNA) is isolated from fresh or FFPE tissues. Total nucleic acid is converted to double-stranded cDNA (dscDNA) and cleaned by solid-phase reversible immobilization (SPRI). Then the dscDNA or gDNA is subjected to end repair, adenylation and ligation with a universal half-functional adapter. The modified DNA sample is then subjected to two rounds of nested low-cycle PCR, where the first round uses target gene-specific primers and the second round uses nested target gene-specific primers that are 5' tagged with a common sequencing adapter. The resulting target amplicon library is cleaned using SPRI and is suitable for use in next generation genomic assays.
Technology
Any of several immunoassay methods that use an enzyme covalently linked to an antigen or antibody as a label; the most common types are enzyme-linked immunosorbent assay (ELISA) and enzyme-multiplied immunoassay technique (EMIT).
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A type of homogenous immunoassay in which sample is applied to a reagent mixture of enzyme-labeled ligand, antibody, and substrate. Substrate binding to enzyme leads to inactivation and unlabeled-ligand in the sample can be quantified by colorimetric analysis.
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An enhancement of immunoelectrophoresis in which a series of post-electrophoretic gel slabs are layered with cellulose-acetate gels saturated with specific antibodies. The resulting antigen-antibody complexes fixed on the second gel may then be stained, allowing sensitive and specific qualitative visual identification of paraproteins by electrophoretic position. It can be used to identify specific proteins in the blood or urine. It has greatest application in the identification and monitoring of monoclonal proteins that are produced in conditions like Waldenstom's macroglobulinaemia and multiple myeloma.
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A type of nucleic acid amplification test in which sample DNA is exposed to a single temperature in the presence of multiple sets of primers and a polymerase with replication and high strand displacement activity. This test negates the need for thermal cyclers and may be a low cost alternative to PCR.
Technology
An imaging technique that uses multiple photons to absorb infrared light, thereby minimizing phototoxicty and photobleaching, making it an ideal method for imaging living specimens, especially those involving deep tissue.
Technology
A method for amplifying a DNA base sequence using multiple rounds of heat denaturation of the DNA and annealing of oligonucleotide primers complementary to flanking regions in the presence of a heat-stable polymerase. This results in duplication of the targeted DNA region. Newly synthesized DNA strands can subsequently serve as additional templates for the same primer sequences, so that successive rounds of primer annealing, strand elongation, and dissociation produce rapid and highly specific amplification of the desired sequence. PCR also can be used to detect the existence of the defined sequence in a DNA sample.