Patent ID: 8383344

Claim:
A method for quantitative determination of a short-length RNA, which has a length of at most 100 nucleotides, comprising the steps of: a) preparing, from a sample comprising said short-length RNA, a template polynucleotide which consists of 1) a single stranded target sequence consisting of the sequence of said short-length RNA, its corresponding DNA sequence or a nucleotide sequence complementary to the sequence of said short-length RNA and 2) a 3′ adjacent nucleotide sequence by appending said 3′ adjacent nucleotide sequence to said single stranded target sequence by nucleotide polymerization; b) hybridizing a single-stranded probe to the 3′ added sequence of the template polynucleotide, wherein said probe comprises a sequence for subsequent amplification of the nucleic acid by polymerase chain reaction (PCR); c) using said template polynucleotide in a reverse transcription or a nucleotide polymerization to obtain a strand of cDNA; d) hybridizing a primer comprising a sequence corresponding to the 5′-end of the template polynucleotide to the cDNA of step (c) double-stranded ouble-stranded target sequence; and e) performing a PCR including as template(s) said cDNA and optionally the template polynucleotide.