C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C07H 21/00 (2006.01) C12N 15/10 (2006.01) C12P 19/34 (2006.01)
Patent
CA 2366231
A method for stimultaneous release and detection of nucleic acids from complex biological samples is described. The invention relates to the combined use of lysis buffers containing strong chaotropic agents such as guanidine thiocyanate to facilitate cell lysis and release of cellular nucleic acids and to the use of a novel type of bicyclic nucleotide analogues, locked nucleic acid (LNA) to detect specific nucleic acids released during lysis by nucleic acid hybridisation. In particular methods are described for the covalent attachment of the catching LNA-oligo. Novel methods for sample preparation of e.g. polyadenylated mRNA species are also presented. The invention further addresses reagents for performing the methods as well as reagents and applications of the method.
L'invention concerne une méthode de libération et de détection simultanée d'acides nucléiques à partir d'échantillons biologiques complexes. L'invention concerne l'utilisation combinée de tampons de lyse contenant des agents chaotropiques forts tels que guanidine thiocyanate afin de faciliter la lyse des cellules et la libération d'acides nucléiques cellulaires, ainsi que l'utilisation d'un nouveau type d'analogues de nucléotides bicycliques, l'acide nucléique bloqué (LNA), afin de détecter des acides nucléiques spécifiques libérés pendant la lyse par l'hybridation des acides nucléiques. En particulier, l'invention a trait à des méthodes de fixation covalente de LNA-oligo de prise. L'invention concerne également de nouvelles méthodes de préparation d'échantillons, par exemple d'espèces d'ARNm polyadénylées. L'invention concerne en outre des réactifs de mise en application des méthodes ainsi que des réactifs et des applications de la méthode.
Exiqon A/s
Ridout & Maybee Llp
LandOfFree
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