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US 8,263,339 B2Granted
Abscription based molecular detection
AssigneeRibomed Biotechnologies, Inc.
InventorsMichelle M. Hanna, David McCarthy
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Abstract
The present invention provides methods for detecting biomarkers based on Abscription®, abortive transcription technology. Particularly, the present invention provides bisulfate free methods for detecting methylation of CpG islands from small samples of DNA. The methods are suitable for multiplexing and can be used to analyze multiple CpG islands from a single sample in a short time.
Claims23
A method for detecting at least one target polynucleotide in a sample comprising:
- a) contacting a sample containing the at least one polynucleotide with a primer pair that specifically hybridizes to and amplifies a target sequence of the at least one polynucleotide, wherein the primer pair consists of:
- i) a first primer comprising
- (1) a 3′ sequence complementary to a first sequence flanking the target sequence of the polynucleotide, and
- (2) a 5′ capture tag;
- and
- ii) a second primer comprising:
- (1) a 3′ sequence complementary to a second sequence flanking the target sequence of the polynucleotide, and
- (2) a 5′ sequence that provides a means for directing Abscription;
- b) amplifying the target sequence from the first and second primers;
- c) contacting the amplified target sequence with an immobilized molecule that binds the 5′ capture tag, thereby capturing the amplified target sequence of the polynucleotide;
- d) transcribing at least one Abscript from the means for directing Abscription;
- and
- e) detecting at least one Abscript transcribed in step d).
10 more elements
Dependencies are inferred from the claim text.