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Reacción en cadena de la polimerasa (PCR)

1983
  • Kary Mullis
Termociclador que amplifica el ADN en un laboratorio de biología molecular.

(Imagen generada únicamente con fines ilustrativos)

La reacción en cadena de la polimerasa (PCR) es una técnica de laboratorio utilizada para amplificar un segmento específico de ADN, generando millones o miles de millones de copias a partir de una pequeña muestra inicial. método Se basa en el ciclo térmico, que consiste en ciclos repetidos de calentamiento y enfriamiento para la fusión del ADN, el apareamiento de los cebadores y la replicación enzimática del ADN mediante una ADN polimerasa termoestable, lo que da como resultado una amplificación exponencial.

The Polymerase Chain Reaction (PCR) is a revolutionary in vitro method for exponentially amplifying a specific target DNA sequence. Its power lies in its ability to generate billions of copies from a minute starting quantity of DNA, making it possible to analyze DNA from samples as small as a single cell. The process is driven by a series of temperature changes, or thermal cycles, managed by a machine called a thermal cycler. Each cycle consists of three core steps. First, Denaturation: the reaction mixture is heated to 94–98 °C for a short period, causing the double-stranded DNA template to separate into single strands. Second, Annealing: the temperature is lowered to 50–65 °C, allowing short, single-stranded DNA primers to bind (anneal) to their complementary sequences on the now single-stranded template DNA. These primers flank the target region to be amplified. Third, Extension: the temperature is raised to 72 °C, the optimal temperature for the key enzyme, a thermostable DNA polymerase, to work. The most commonly used is Taq polymerase, isolated from the thermophilic bacterium *Thermus aquaticus*. The polymerase binds to the primer-template complex and begins synthesizing a new complementary DNA strand, extending from the primer. This three-step cycle is repeated 20-40 times. With each cycle, the number of copies of the target DNA sequence doubles, leading to an exponential amplification, mathematically described as [latex]2^n[/latex], where n is the number of cycles. The result is a solution containing a vast quantity of the specific DNA fragment, which can then be easily visualized by gel electrophoresis or used in subsequent biología molecular aplicaciones como la secuenciación o la clonación.

UNESCO Nomenclature: 2406
- Biología molecular

Tipo

Proceso químico

Ruptura

Revolucionario

Uso

Uso generalizado

Precursores

  • discovery of DNA structure
  • understanding of DNA replication principles
  • isolation of DNA polymerase enzymes
  • discovery of thermostable organisms and their enzymes (e.g., Taq polymerase)
  • development of oligonucleotide synthesis for primers

Aplicaciones

  • DNA fingerprinting for forensics
  • diagnósticos médicos para enfermedades infecciosas (por ejemplo, COVID-19)
  • genetic testing for hereditary diseases
  • cloning genes
  • sequencing genomes
  • phylogenetic analysis

Patentes:

  • US4683195
  • US4683202
  • US4965188

Ideas para posibles innovaciones

  • portable, real-time PCR devices for point-of-care disease diagnosis
  • digital PCR (dPCR) for absolute quantification of nucleic acids
  • multiplex PCR for simultaneous detection of multiple pathogens
  • reverse transcription PCR (RT-PCR) for RNA analysis and gene expression studies
  • in-situ PCR for localizing DNA/RNA within cells or tissues
  • high-fidelity PCR enzymes for reducing amplification errors in sequencing
  • rapid PCR protocols that complete amplification in minutes
  • integration of PCR with microfluidos ‘lab-on-a-chip’ systems
  • using PCR for environmental DNA (eDNA) monitoring of biodiversidad
  • PCR-based assays for food authenticity and GMO detection
Relacionado con: PCR, reacción en cadena de la polimerasa, amplificación del ADN, kary mullis, taq polimerasa, termociclado, medicina forense, diagnóstico, clonación molecular, pruebas genéticas.

Contexto histórico

Reacción en cadena de la polimerasa (PCR)

1973
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1983
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1990
1970
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1983
1987
1990
1990
1990

(Si la fecha es desconocida o no es relevante, por ejemplo "mecánica de fluidos", se proporciona una estimación redondeada de su aparición notable)

décimo año aniversario

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