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, Annexe 5 : protocole d'extraction d'ARN
, Ajouter 450µL de tampon RLC + 4,5µL de ?-mercaptoéthanol et vortexer
, Ajouter 0,5V d'éthanol ARN free et agiter par retournement doucement
, Laver la membrane avec 350µL de RW1, centrifugation 30 secondes à 10 000rpm
Ajouter 80µL d'un mélange DNase + tampon ,
, Ajouter 350µL de RW1, centrifugation 30 secondes à 10 000rpm
, Vider la partie liquide
, Ajouter 500µL de RPE et centrifuger 30 secondes à 10 000rpm
, Vider la partie liquide et remettre 350µL de RPE + centrifugation 30 secondes à, vol.10, p.0
, Enlever le tube de récupération, et le remplacer par un eppendorf ARN free
, Centrifuger 1 minute pour récupérer l'ARN dans l'eppendorf
, Ajouter 200µL de LiCl 4M (permet la dégradation de potentielle RNases), retourner doucement 20) Stocker à-20°C