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25053641
RT PCR METHOD
Description
biologia molecular
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biologia
Flowchart by
Manuel Enrique Alavrez
, updated more than 1 year ago
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Created by
Manuel Enrique Alavrez
over 4 years ago
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Metodología a seguir para detección de ARNm de VPH
1.-Se toman muestras del cervix y se almacenaron a (-80ºC)
2.- El ARN de todas las células se preparó usando Trizol (Life Technologies, Carlsbad, CA) d
3.- Se preparan ADNc (complementario) en sistema de síntesis de primera cadena SuperScript III para RT-PCR
4.- Uso de PCR semicuantitativa para amplificar simultáneamente el ADNc celular de GAPDH y HPV L1.
5.-Analisis de muestras
Uso de centrifugación para sedimentar ARN
Analisis de pureza por AGILENT 2100
Rtpcr Cebadores (binary/octet-stream)
síntesis de ADNc se realizó en ácido nucleico extraído tratado con DNase-treated
TRANSCRIPCIÓN INVERSA
Se multiplicaron las cadenas de ADNc
REPLICACIÓN DE ADNc
Se usaron cebadores y primers para determinar la cantidad en tiempo real de carga viral (ARNm)
Rtpcr Dogma (binary/octet-stream)
Captura1 (binary/octet-stream)
Captura2 (binary/octet-stream)
Captura4 (binary/octet-stream)
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