Real-time nucleic acid detection processes and compositions

摘要

This invention provides compositions for use in real time nucleic acid detection processes. Such real time nucleic acid detection processes are carried out with energy transfer elements attached to nucleic acid primers, nucleotides, nucleic acid probes or nucleic acid binding agents. Real time nucleic acid detection allows for the qualitative or quantitative detection or determination of single-stranded or double-stranded nucleic acids of interest in a sample. Other processes are provided by this invention including processes for removing a portion of a homopolymeric sequence, e.g., poly A sequence or tail, from an analyte or library of analytes. Compositions useful in carrying out such removal processes are also described and provided.

主权项

1. A composition of matter comprising at least two parts:
a first part comprising at least two segments, wherein said first segment comprises a first nucleic acid primer and at least one first energy transfer element, wherein said first energy transfer element is: (a) an energy donor and (b) covalently attached directly or via a linker arm to said first nucleic acid primer and said second segment comprises a primer-extended nucleic sequence complementary to at least a portion of a nucleic acid target sequence; and a second part comprising at least two segments, wherein said first segment of said second part comprises a second nucleic acid primer and at least one second energy transfer element, wherein said second energy transfer element is: (a) an energy acceptor and (b) covalently attached directly or via a linker arm to said first nucleic acid primer; and wherein said second segment of said second part comprises a primer-extended nucleic acid sequence which is identical to at least a portion of said nucleic acid target sequence; wherein said first nucleic acid primer does not comprise said second energy transfer element, and wherein said second nucleic acid primer does not comprise said first nucleic acid energy element; wherein said first primer is not complementary to said second primer or nucleic acid in the absence of a target dependent extension step; and wherein neither of said nucleic acid primers is fixed or immobilized to a solid support.