File:Sanger-sequencing.svg

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English: The Sanger (chain-termination) method for DNA sequencing. (1) A primer is annealed to a sequence, (2) Reagents are added to the primer and template, including: DNA polymerase, dNTPs, and a small amount of all four dideoxynucleotides (ddNTPs) labeled with fluorophores. During primer elongation, the random insertion of a ddNTP instead of a dNTP terminates synthesis of the chain because DNA polymerase cannot react with the missing hydroxyl. This produces all possible lengths of chains. (3) The products are separated on a single lane capillary gel, where the resulting bands are read by a imaging system. (4) This produces several hundred thousand nucleotides a day, data which require storage and subsequent computational analysis
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Español: El método de terminación de la cadena de Sanger. (1) Un cebador se alínea a la secuencia, (2) Se añaden reactivos al cebador y ADN molde incluyendo: polimerasa de ADN, dNTPs, una pequeña cantidad de todos los cuatro didesoxinucleótidos (ddNTPs) marcados con fluoróforos. Durante la elongación de los cebadores, una inserción aleatoria de un ddNTP en vez de un dNTP termina la síntesis de la cadena debido a que la ADN polimerasa no puede reaccionar con el hidroxilo faltante. Esto produce todas las posibles longitudes de las cadenas. (3) Los productos se separan en un gel capilar, donde las bandas resultantes se leen por un sistema de imágenes. (4) Esto produce muchos cientos de miles de nucleótidos al día, y los datos requieren almacenaje y posterior análisis por computadora.
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Author Estevezj

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Date/TimeThumbnailDimensionsUserComment
current19:55, 20 December 2012Thumbnail for version as of 19:55, 20 December 2012673 × 463 (2.67 MB)Estevezj (talk | contribs)included chemical structures for ddNTPs and dNTPs
07:29, 20 December 2012Thumbnail for version as of 07:29, 20 December 2012673 × 463 (2.63 MB)Estevezj (talk | contribs)Replaced Sans with Helvetica, deleted wonky layer.
06:52, 20 December 2012Thumbnail for version as of 06:52, 20 December 2012650 × 387 (2.3 MB)Estevezj (talk | contribs)Reverted to version as of 06:50, 20 December 2012 bad image
06:51, 20 December 2012Thumbnail for version as of 06:51, 20 December 2012671 × 440 (2.64 MB)Estevezj (talk | contribs)English labels.
06:50, 20 December 2012Thumbnail for version as of 06:50, 20 December 2012650 × 387 (2.3 MB)Estevezj (talk | contribs)User created page with UploadWizard
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