METHODS FOR ENHANCING NUCLEIC ACID HYBRIDIZATION

摘要

A composition, comprising a first oligonucleotide having the structure 5'-Y1-L1-X-L2_2-Y2-3' Y1 comprises a sequence of four or more DNA or RNA nucleotides, including a first nucleotide N1 having a 3' phosphate covalently linked to L1. Y2 comprises a sequence of four or more DNA or RNA nucleotides, including a second nucleotide N2 having a 5' phosphate covalently linked to L2. L1and L2 each independently are a direct bond or a C1 C7 alkyl, alkynyl, alkenyl, heteroalkyl, substituted alkyl, aryl, heteroaryl, substituted aryl, cycloalkyl, alkylaryl, or alkoxyl group. X is R/ is a hydrogen or a d-Cg alkyl. M is a quencher. The first oligonucleotide is adapted to hybridize to a second oligonucleotide having the structure 3'-?3-?4-5'. Y3 comprises a sequence of four or more DNA or RNA nucleotides, including a third nucleotide N3. Y4 comprises a sequence of four or more DNA or RNA nucleotides, including a fourth nucleotide N4 that is directly attached to nucleotide N3. If the first oligonucleotide hybridizes to the second oligonucleotide, Ni base pairs with N3 and N2 base pairs with N4.to form a duplex having a Tm that is greater than the Tm of a duplex formed between the second oligonucleotide and a third oligonucleotide having the structure 5'-Y1-Y2-3'.