Reacción en Cadena de la Polimerasa (PCR)

Descripción General

Definición

La reacción en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum cadena de la polimerasa ( PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR), por sus siglas en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum inglés) es una técnica que amplifica secuencias de ADN específicas in vitro y de manera exponencial y se utiliza para el diagnóstico, la identificación de virus Virus Viruses are infectious, obligate intracellular parasites composed of a nucleic acid core surrounded by a protein capsid. Viruses can be either naked (non-enveloped) or enveloped. The classification of viruses is complex and based on many factors, including type and structure of the nucleoid and capsid, the presence of an envelope, the replication cycle, and the host range. Virology y el análisis.

Componentes necesarios

• ADN molde (muestra con la secuencia genómica)
• Cebadores:
  • Oligonucleótidos cuidadosamente diseñados para una secuencia diana específica
  • Complementarios al AL Amyloidosis extremo 3′ de la región diana
• Desoxinucleósido trifosfato (dNTP)
• ADN polimerasa:
  • Debe ser termoestable para sobrevivir varias rondas de calentamiento/enfriamento
  • Realiza la replicación

Procedimiento

Para amplificar el ADN o el ARN, las siguientes etapas se repiten en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum ciclos (a menudo hasta 30–40 veces):

• Desnaturalización
• Hibridación de los LOS Neisseria cebadores
• Síntesis

Desnaturalización

• La muestra se calienta a aproximadamente 95°C (aproximadamente 203°F).
• Los LOS Neisseria enlaces de hidrógeno se rompen → separación del ADN en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum hebras individuales

Hibridación de los LOS Neisseria cebadores

• La temperatura se reduce a aproximadamente 50–65°C (aproximadamente 122–149°F).
• Los LOS Neisseria cebadores se unen al AL Amyloidosis extremo 3′ de las hebras de ADN molde.
• Sigue la unión de la ADN polimerasa.

Síntesis/elongación

• La temperatura se aumenta a aproximadamente 70–72°C (aproximadamente 158–162°F).
• La ADN polimerasa se activa → agrega nucleótidos libres al AL Amyloidosis extremo 3′ de los LOS Neisseria cebadores
• Crea una nueva hebra de ADN idéntica al AL Amyloidosis molde original

Repetición de los LOS Neisseria ciclos

• El ADN recién creado sirve como molde para otros ciclos de PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR).
• Se replica el ADN exponencialmente en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum un factor 2^x, donde x es el número de ciclos de replicación
• El segmento de ADN diana se puede amplificar 10⁵–10⁶ veces.

Análisis

• Luego, el segmento de ADN amplificado se puede observar con técnicas como la electroforesis en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum gel.
• Los LOS Neisseria resultados deben compararse con ambos:
  • Control positivo
  • Control negativo

Variaciones

Hay muchas variaciones de la técnica básica de PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR). Algunos de los LOS Neisseria más comunes incluyen:

• PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) de transcripción inversa
• PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) cuantitativa

PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) de transcripción inversa

• Utiliza ARN como molde
• Usa transcriptasa inversa → crea una hebra complementaria de ADN (híbrido ADN/ARN)
• La porción de ARN es degradada por la transcriptasa inversa o ribonucleasa H.
• La ADN polimerasa agrega nucleótidos complementarios a la hebra de ADN → molécula de ADN completa
• El nuevo ADN se convierte en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum el molde → pasa por el proceso normal de PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) desde allí

PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) cuantitativa

• También llamado PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum tiempo real
• Supervisa la amplificación agregando:
  • Sonda fluorescente (secuencia específica)
  • Colorante fluorescente (se intercala con el ADN)
• Permite detección y visualización con software especializado
• Útil para:
  • Cuantificación (e.g., carga viral)
  • Detección rápida de una secuencia específica (sin necesidad de técnicas de análisis posteriores a la PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR))

Usos

Medicina forense

• La PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) permite la comparación de pequeñas muestras de evidencia de ADN de la escena del crimen con el ADN de los LOS Neisseria sospechosos.
• Prueba de paternidad

Genotipificación y secuenciación

• Identificación de mutaciones genéticas:
  • Permite el diagnóstico de muchas enfermedades genéticas
  • Se puede realizar en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum células embrionarias para determinar si un embrión tiene predisposición a ciertas afecciones
  • Puede identificar portadores genéticos → útil para el asesoramiento genético
• El reconocimiento de oncogenes Oncogenes Genes whose gain-of-function alterations lead to neoplastic cell transformation. They include, for example, genes for activators or stimulators of cell proliferation such as growth factors, growth factor receptors, protein kinases, signal transducers, nuclear phosphoproteins, and transcription factors. A prefix of ‘v-‘ before oncogene symbols indicates oncogenes captured and transmitted by retroviruses; the prefix ‘c-‘ before the gene symbol of an oncogene indicates it is the cellular homolog (proto-oncogenes) of a v-oncogene. Carcinogenesis alterados puede permitir una terapia dirigida contra el cáncer.
• Se puede utilizar para la tipificación de tejidos en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum la implantación de órganos

Enfermedad infecciosa

La PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) ha HA Hemolytic anemia (HA) is the term given to a large group of anemias that are caused by the premature destruction/hemolysis of circulating red blood cells (RBCs). Hemolysis can occur within (intravascular hemolysis) or outside the blood vessels (extravascular hemolysis). Hemolytic Anemia mejorado significativamente el potencial diagnóstico de enfermedades infecciosas.

• Puede determinar rápidamente la identidad de microbios que tradicionalmente eran:
  • De crecimiento lento en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum cultivo
  • Imposibles de cultivar
• Ejemplos:
  • VIH
  • Virus Virus Viruses are infectious, obligate intracellular parasites composed of a nucleic acid core surrounded by a protein capsid. Viruses can be either naked (non-enveloped) or enveloped. The classification of viruses is complex and based on many factors, including type and structure of the nucleoid and capsid, the presence of an envelope, the replication cycle, and the host range. Virology herpes simple
  • SARS-CoV-2
  • Influenza Influenza Influenza viruses are members of the Orthomyxoviridae family and the causative organisms of influenza, a highly contagious febrile respiratory disease. There are 3 primary influenza viruses (A, B, and C) and various subtypes, which are classified based on their virulent surface antigens, hemagglutinin (HA) and neuraminidase (NA). Influenza typically presents with a fever, myalgia, headache, and symptoms of an upper respiratory infection. Influenza Viruses/Influenza
  • Mycobacterium Mycobacterium Mycobacterium is a genus of the family Mycobacteriaceae in the phylum Actinobacteria. Mycobacteria comprise more than 150 species of facultative intracellular bacilli that are mostly obligate aerobes. Mycobacteria are responsible for multiple human infections including serious diseases, such as tuberculosis (M. tuberculosis), leprosy (M. leprae), and M. avium complex infections. Mycobacterium
  • Borrelia burgdorferi Borrelia burgdorferi A specific species of bacteria, part of the borrelia burgdorferi group, whose common name is lyme disease spirochete. Borrelia
• La PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) cuantitativa permite la cuantificación de la carga microbiana (además del análisis cualitativo).
• También se puede utilizar para detectar genes Genes A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms. DNA Types and Structure que codifican la resistencia a los LOS Neisseria antibióticos

Ventajas y Desventajas

Ventajas

• Rápida
• Bajo costo
• Relativamente fácil de usar
• Altamente específica
• Sensible (permite su uso en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum tamaños de muestra muy pequeños)
• Puede amplificar el ADN exponencialmente para crear un número casi infinito de copias del ADN diana
• La máquina de PCR PCR Polymerase chain reaction (PCR) is a technique that amplifies DNA fragments exponentially for analysis. The process is highly specific, allowing for the targeting of specific genomic sequences, even with minuscule sample amounts. The PCR cycles multiple times through 3 phases: denaturation of the template DNA, annealing of a specific primer to the individual DNA strands, and synthesis/elongation of new DNA molecules. Polymerase Chain Reaction (PCR) replica el ADN in vitro:
  • No se requieren bacterias
  • Sin extracción de ADN

Desventajas

• Extremadamente sensible → propenso a errores por contaminación de ARN o ADN
• Los LOS Neisseria cebadores requieren datos de secuencia (solo se pueden usar para identificar la presencia de un gen conocido).
• Potencial de los LOS Neisseria cebadores para hibridarse con otras secuencias similares → amplificación de la secuencia genética incorrecta
• Formación de dímeros de cebadores:
  • Los LOS Neisseria cebadores se hibridan entre sí a través de bases Bases Usually a hydroxide of lithium, sodium, potassium, rubidium or cesium, but also the carbonates of these metals, ammonia, and the amines. Acid-Base Balance complementarias.
  • Da como resultado la amplificación del dímero en EN Erythema nodosum is an immune-mediated panniculitis (inflammation of the subcutaneous fat) caused by a type IV (delayed-type) hypersensitivity reaction. It commonly manifests in young women as tender, erythematous nodules on the shins. Erythema Nodosum lugar de la secuencia diana