Generic method for the stabilization of specific RNA
The present invention relates to methods and compositions for the stabilization of specific RNA molecules that can either be the target for detection or the control standard.
1. A method of preventing or reducing degradation of a segment of a single-stranded RNA template that is amplified in an amplification reaction, the method comprising the steps of:
a) providing the single-stranded RNA template;
b) hybridizing the segment of the single-stranded RNA template with one or more oligonucleotides whose sequences are completely or partially complementary to the segment of the single-stranded RNA template that is amplified; and
c) reverse transcribing and amplifying the segment of the single-stranded RNA template under reaction conditions whereby the one or more oligonudeotides do not interfere with reverse transcription and amplification and whereby each one oligonucleotide from the one or more oligonucleotides is characterized by both being between 11 nucleotides and 50 nucleotides in length and having a melting temperature that is at least 5° C. lower than an extension temperature used during amplification, and wherein the sequence of each one oligonudeotide from the one or more oligonudeotides does not overlap with the sequence of another oligonudeotide from the one or more oligonudeotides; and
the one or more oligonucleotides are present at a concentration that is at least fifty-fold lower than concentrations of primers and probes used during reverse transcription and amplification;
wherein the one or more oligonudeotides comprise a group of oligonudeotides whose sequences consist of SEQ ID NOs: 1-10.