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US 6,670,122 B2Granted
Method for detecting transcription templates
AssigneeAffymetrix, Inc.
InventorsCarsten Rosenow, Rini Saxena
Life of this patent
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Abstract
Methods are provided for detecting the sense and antisense transcripts and for determining template strand of a genomic DNA. Exemplary methods include reverse transcribing transcripts without second strand synthesis. The resulting single stranded DNA is labeled and detected using nucleic acid probe arrays. In a particularly embodiment, actinomycin is used to inhibit the synthesis of second strand cDNA during reverse transcription.
Claims25
A method for detecting a plurality of transcripts comprising:
- synthesizing a plurality of cDNAs complementary with the transcripts by reverse transcription;
- wherein the synthesis of second strand cDNA is inhibited;
- and
- hybridizing the cDNAs or nucleic acids derived from the cDNAs with a nucleic acid probe array to detect the transcripts.
1 more element
A method for detecting transcribed regions of a genome comprising
- obtaining a sample comprising transcripts transcribed from the genome;
- synthesizing single stranded cDNAs complementary with the transcripts, wherein the synthesis of second strand cDNA is inhibited;
- and
- hybridizing the cDNAs or nucleic acids derived from the cDNAs with a nucleic acid probe array, wherein the nucleic acid probe array has probes targeting both strands of the genomic DNA in interested regions.
1 more element
An assay kit comprising:
- reagents necessary for a reverse transcription reaction;
- an inhibitor of second strand cDNA synthesis;
- and
- a nucleic acid probe array.
1 more element
Dependencies are inferred from the claim text.