Methods and compositions for amplification of rna sequences

Details Methods and compositions for amplification of rna sequences Methods and compositions for amplification of rna sequences

C - Chemistry, Metallurgy

12

P

C12P 19/34 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2439074

The invention provides methods for linear and exponential amplification of RNA. They are particularly suitable for amplifying a plurality of RNA species in a sample. The methods are based on hybridization of polynucleotide comprising a propromoter sequence to a primer extension product to generate an intermediate polynucleotide capable of driving transcription, whereby multiple copies of RNA products comprising sequences complementary to an RNA sequence of interest are generated. The methods are useful for preparation of nucleic acid libraries and substrates for analysis of gene expression of cells in biological samples. The invention also provides compositions and kits for practicing the amplification methods, as well as methods which use the amplification products.

Cette invention concerne des méthodes d'amplification linéaire et exponentielle d'ARN. Ces méthodes conviennent tout particulièrement pour l'amplification d'une pluralité de types d'ARN dans un échantillon. Elles reposent sur un hybridation d'un polynucléotide comprenant une séquence promoteur avec un produit de rallonge d'amorce visant à générer un polynucléotide intermédiaire capable de commander une transcription, avec production de copies multiples de produits d'ARN comprenant des séquences complémentaires de la séquence d'ARN d'intérêt. Ces méthodes conviennent pour l'établissement de bibliothèques et de substrats d'acides nucléiques en vue de l'analyse de l'expression génique de cellules dans des échantillons biologiques. De plus, l'invention concerne des compositions et des trousses permettant de mettre en oeuvre des méthodes d'amplification ainsi que des méthodes recourant à des produits d'amplification.

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