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PCR01:32

PCR

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Overview
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The Replisome03:01

The Replisome

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DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
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Maxam-Gilbert Sequencing01:05

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In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
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Translesion DNA Polymerases02:10

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
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DNA Replication02:40

DNA Replication

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DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication...
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DNA as a Genetic Template02:05

DNA as a Genetic Template

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Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...
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Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
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Síntesis de ADN en varios pasos con plantilla utilizando una plantilla universal.

Yizhou Li1, Peng Zhao, Mingda Zhang

  • 1Key Laboratory of Bioorganic Chemistry and Molecular Engineering of the Ministry of Education, Beijing National Laboratory of Molecular Sciences (BNLMS), College of Chemistry and Molecular Engineering, Peking University , 202 Chengfu Road, Beijing, China 100871.

Journal of the American Chemical Society
|November 16, 2013
PubMed
Resumen
Este resumen es generado por máquina.

Un nuevo método de síntesis con plantilla de ADN utiliza una plantilla de ADN universal para construir diversas bibliotecas codificadas con ADN. Este enfoque simplifica la síntesis de bibliotecas, la selección de objetivos y la decodificación de golpes para aplicaciones químicas más amplias.

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Área de la Ciencia:

  • Biología Química Biología química.
  • Biología Molecular Biología Molecular
  • Química sintética de la química sintética.

Sus antecedentes:

  • Las bibliotecas codificadas por ADN (DEL) ofrecen una poderosa plataforma para el descubrimiento de fármacos.
  • Los métodos de síntesis DEL actuales pueden ser complejos y requieren múltiples plantillas de ADN.

Objetivo del estudio:

  • Desarrollar un método de síntesis simplificado con plantillas de ADN para crear diversas bibliotecas codificadas con ADN.
  • Demostrar la utilidad de una nueva "plantilla universal" para la construcción y el análisis eficientes de bibliotecas.

Principales métodos:

  • Se diseñó una sola plantilla de ADN que incorporaba deoxinosina para el emparejamiento indiscriminado de bases.
  • Se integraron en el sistema los enlaces fotocéptiles y la codificación directa por el ADN reactivo.
  • La plantilla universal se utilizó para la síntesis de bibliotecas, la selección de objetivos y la decodificación de golpes.

Principales resultados:

  • Construcción exitosa de una biblioteca completa codificada con ADN utilizando una sola plantilla de ADN universal.
  • Eficiencia demostrada en la dirección de múltiples reacciones químicas con variados ADNs reactivos.
  • Capacidades validadas en la síntesis de bibliotecas, selección de objetivos y decodificación de golpes.

Conclusiones:

  • El método de plantilla universal proporciona un enfoque general y sencillo para preparar diversas bibliotecas codificadas con ADN.
  • Esta técnica simplifica la síntesis DEL, permitiendo aplicaciones más amplias en biología química y descubrimiento de fármacos.