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PCR

PCR

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction

Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Apta-PCR.

Alessandro Pinto^1,2, Pedro Nadal Polo^1,3, Miriam Jauest Rubio¹

¹Department of Chemical Engineering, Universitat Rovira I Virgili, Avinguda Päısos Catalans 26, Tarragona, 43007, Spain.

Methods in Molecular Biology (Clifton, N.J.)

|November 11, 2015

Summary

This summary is machine-generated.

Real-time Apta-PCR combines nucleic acid amplification with aptamers for sensitive analyte detection. This adaptable method improves detection limits for applications in food safety and clinical diagnostics.

Keywords:

Apta-PCR Aptamer Protein quantification

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Area of Science:

Biotechnology
Molecular Diagnostics
Analytical Chemistry

Background:

Real-time Apta-PCR integrates nucleic acid amplification with aptamer selectivity.
This technique offers high sensitivity for detecting target analytes.
Existing methods may have limitations in detection limits for certain targets.

Purpose of the Study:

To describe a generic real-time Apta-PCR methodology.
To demonstrate its adaptability for various targets.
To validate the method using a model target, β-conglutin.

Main Methods:

Development of a generic real-time Apta-PCR protocol.
Utilizing a competitive assay format.
Selection and application of aptamers for target recognition.

Main Results:

The real-time Apta-PCR methodology was successfully established.
Demonstrated potential for ultra-low detection limits.
Showcased adaptability for detecting a model target (β-conglutin).

Conclusions:

Real-time Apta-PCR is a versatile platform for sensitive analyte detection.
The method can be adapted for diverse applications, including food quality and clinical diagnostics.
Improved detection limits are achievable, even with lower-affinity aptamers.