Methods for improving rna transcription reactions

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01) C07H 19/00 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2487660

Methods are described for eliminating single-stranded oligonucleotides from a sample prior to RNA transcription, thereby reducing non-template derived production of RNA. In one embodiment, a sample containing the template for RNA transcription is treated with one or more exonucleases to remove single- stranded oligonucleotides from the reaction mixture prior to RNA transcription. In another embodiment, the sample containing the template for RNA transcription is contacted with an oligonucleotide complementary to the single-stranded oligonucleotide present in the sample, and allowed to hybridize to form double-stranded oligonucleotides.

L'invention concerne des procédés permettant d'éliminer des oligonucléotides monocaténaires contenus dans un échantillon avant une transcription d'ARN, et par conséquent de réduire la production d'ARN sans l'aide de matrices. Dans un mode de réalisation de cette invention, un échantillon contenant la matrice de transcription d'ARN est traitée au moyen d'une ou de plusieurs exonucléases pour supprimer les oligonucléotides monocaténaires contenus dans le mélange réactionnel avant une transcription d'ARN. Dans un autre mode de réalisation, l'échantillon renfermant la matrice de transcription d'ARN est mise en contact avec un oligonucléotide qui est complémentaire à l'oligonucléotide monocaténaire présent dans l'échantillon et l'hybridation entre cet oligonucléotide complémentaire et l'oligonucléotide monocaténaire est permise pour former des oligonucléotides bicaténaires.

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