C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C12N 15/09 (2006.01) C12P 19/34 (2006.01)
Patent
CA 2437737
The present invention pertains to a method that will increase the efficiency of second strand cDNA synthesis through a mechanism of "terminal continuation" before further RNA amplification by RNA transcription using, for example, a bacteriophage promoter. In a specific embodiment, a transcription promoter is attached to the 5' region of cDNA through the same mechanism of "terminal continuation". Genetic signals are subsequently amplified in a linear manner through RNA transcription. In specific embodiments, the orientation of the transcribed RNA is either sense or antisense, depending on the desired downstream application. In other embodiments, the present invention pertains to methods for extraction and amplification of RNA, particularly mRNA, from histologically stained tissues and cells.
L'invention porte sur un procédé améliorant l'efficacité de la synthèse d'ADNc de deuxième brin recourant à un mécanisme de "prolongation de terminal" avant d'effectuer une amplification d'ARN par transcription utilisant par exemple un promoteur bactériophage. Dans une exécution spécifique, un promoteur de transcription est lié à la région 5' de l'ADNc à l'aide du susdit mécanisme. Les signaux génétiques sont ensuite amplifiés de manière linéaire par transcription d'ARN. Dans d'autres exécutions spécifiques l'orientation de l'ARN transcrit est du type sens ou antisens selon l'application aval désirée. L'invention à par ailleurs trait à des procédés d'extraction et d'amplification d'ARN particulièrement d'ARNm provenant de tissus et de cellules colorés histologiquement.
Che Shaoli
Ginsberg Stephen D.
Macrae & Co.
Research Foundation For Mental Hygiene Inc.
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