Nucleic acid labeling methods

Details Nucleic acid labeling methods Nucleic acid labeling methods

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07H 19/04 (2006.01) C07H 19/073 (2006.01) C07H 19/10 (2006.01) C07H 19/173 (2006.01) C07H 19/20 (2006.01) C07H 21/00 (2006.01) C12N 15/11 (2006.01) C12P 19/34 (2006.01) C40B 30/04 (2006.01) C40B 40/06 (2006.01) G01N 33/532 (2006.01)

Patent

CA 2525358

In one aspect of the invention, a method is provided for end-labeling RNA (total RNA, mRNA, cRNA or fragmented RNA). In one aspect of the present invention, T4 RNA ligase is used to attach a 3'-labeled AMP or CMP donor to an RNA acceptor molecule. In another embodiment, a pyrophosphate molecule 3'-AppN-3'-linker- detectable moiety is used as donor molecule. In another aspect of the present invention, a method of detecting the presence of an RNA of interest in a sample is provided, the method having the following steps: providing the sample comprising RNA which may or may not have said RNA of interest; treating the sample with a fragmenting reagent to provide RNA fragments; removing phosphate groups from said fragments to provide fragments with free 3' OH groups; ligating said fragment with a labeling reagent according to the instant invention; providing a nucleic acid array having probes directed to said RNA of interest; hybridizing the labeled nucleic acid fragments to said nucleic acid array; and determining the extent of hybridization to said probes to determine the presence of said RNA of interest.

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