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US 8,802,867 B2Granted
Method for producing a sense RNA molecule
AssigneeGenisphere, LLC
InventorsRobert C. Getts, Jaime Boyle
Life of this patent
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Abstract
Methods and kits are provided for producing sense RNA molecules. The sense RNA molecules can be used in various research and diagnostic applications, such as gene expression studies involving nucleic acid microarrays.
Claims32
A method for producing a sense RNA molecule, comprising:
- a) providing a single stranded cDNA molecule having 5′ and 3′ ends;
- b) attaching a homopolymeric tail for annealing a single stranded promoter template to the 3′ end of said single stranded cDNA molecule using terminal deoxynucleotidyl transferase;
- c) annealing a single stranded promoter template consisting essentially of a single RNA polymerase promoter sequence at a 5′ end and a sequence complementary to the homopolymeric tail at a 3′ end thereof to said homopolymeric tail through base pairing with the complementary sequence of the single stranded promoter template, wherein said single stranded promoter template is not extendable with DNA polymerase;
- d) extending said homopolymeric tail such that said single stranded promoter template is converted to a double stranded promoter;
- and
- e) initiating RNA transcription using an RNA polymerase which recognizes said double stranded promoter,
- thereby producing a sense RNA (sRNA) molecule.
4 more elements
Dependencies are inferred from the claim text.