Anchored Multiplex Pcr
Anchored Multiplex Pcr - Related Videos
10:15
Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)
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Cited by 21 •
2011
An inexpensive, high throughput method for simultaneous detection of up to 43 molecular targets is described. Applications of mPCR/RLB include microbial typing and detection of multiple pathogens from clinical samples.
03:41
Agarose Gel Electrophoresis of DNA Amplicons Post PCR: A Method to Analyze Products of Multiplex PCR and Evaluate PCR Reaction Success
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2025
In this video, we demonstrate the separation of bacterial PCR-amplified DNA using agarose gel electrophoresis. Agarose gel functions as a molecular sieve, enabling the negatively-charged DNA to migrate based upon their size under an applied electric field.
07:27
High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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Cited by 13 •
2011
Here we describe an optimized multiplex reverse transcriptase quantitative PCR (qRT-PCR) protocol in combination with a microfluidic platform as a cost and time effective high-throughput screening tool for microRNA (miRNA) expression levels, especially when working with limited amounts of sample.
09:03
Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
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Cited by 18 •
2013
We demonstrate a simple multiplex PCR assay for quick-screening and typing of Staphylococcal Cassette Chromosome mec (SCCmec) types I-V for methicillin-resistant Staphylococcus aureus, and provide some of the vital steps and procedural nuances that make it successful for adapting this assay to individual laboratories.
01:25
The Anchoring-and-Adjustment Heuristic
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2020
In order to make good decisions, we use our knowledge and our reasoning. Often, this knowledge and reasoning is sound and solid. However, sometimes, we are swayed by biases or by others manipulating a situation. For example, let’s say you and three friends wanted to rent a house and had a combined target budget of $1,600. The realtor shows you only very run-down houses for $1,600 and then shows you a very nice house for $2,000. Might you ask each person to pay more in rent to get the $2,000...
FAQs
A gene-specific primer targets the known portion of a rearranged genomic sequence or transcript, while a universal primer binds an adapter-linked or otherwise known sequence on the other side. Because the second primer does not require a predefined partner sequence, amplification can proceed even when that fusion partner was not anticipated, supporting broader detection of gene rearrangements.
Gene-specific primers determine which genes or sequence regions are selected for amplification. Universal primers recognize the shared adapter-derived or otherwise known sequence and provide a common amplification partner across targets. This division of roles allows one assay design to interrogate multiple candidate genes while retaining the ability to recover rearrangements involving different, previously unspecified partners.
Multiplexing allows many targets to be amplified in parallel rather than processing each gene alteration separately. When combined with next-generation sequencing, this design supports simultaneous assessment of rearrangements, mutations, and other sequence alterations from the same assay. The result can be broader molecular profiling while making efficient use of limited tumor material.
The workflow begins with DNA or RNA-derived complementary DNA, followed by preparation that provides an adapter-ligated or otherwise known sequence for universal-primer binding. Gene-specific and universal primers then amplify selected regions in parallel, and the resulting targeted products can be analyzed with next-generation sequencing. This sequence connects sample preparation, multiplex amplification, and molecular interpretation.
Anchored multiplex PCR can be performed using DNA or RNA-derived complementary DNA as the starting molecular material. That flexibility allows the assay to address sequence alterations present in genomic DNA as well as information represented in RNA transcripts after conversion to cDNA. The selected input therefore depends on which molecular form is available from the tissue and relevant to the analysis.
The method is particularly useful when tumor tissue is limited or when the possible fusion partner is not known in advance. Its broad partner detection can contribute to molecular diagnosis, tumor classification, and biomarker discovery. By identifying oncogenic fusions and actionable variants, the resulting profile can also inform selection of targeted therapies.