Quantization Procedures microRNA and small interfering RNA

摘要

A method for quantitative determination of a short-length RNA, which has a length of at most 100 nucleotides, comprising a) preparing, from a sample comprising said short-length RNA, a polynucleotide mold consisting of 1 ) a single stranded target sequence consisting of the sequence of said short-length RNA, its corresponding DNA sequence or nucleotide sequence complementary to the sequence of said RNA short and two long) a nucleotide sequence adjacent 5 'and / or 3 ', wherein said nucleotide sequence adjacent 5' and / or 3 'is a polynucleotide consisting of identical nucleotides, b) using said polynucleotide template in a reverse transcription or nucleotide polymerization to obtain a strand cDNA c) performing PCR quantitative real time (qPCR) including as said cDNA mold or molds and, optionally, the polynucleotide template, wherein 1) the primers used for qPCR in step c are selected from - at least 2 oligonucleotides, wherein at least one of said oligonucleotides corresponds to or is complementary to a sequence in the nucleotide sequence adjacent 5 'or 3' and - at least 2 oligonucleotides, the at least one of said oligonucleotides corresponds to or is complementary to a contiguous sequence in the polynucleotide mold constituted by the single stranded target sequence and part of the nucleotide sequence 5 'or 3' adjacent, or wherein 2) the reaction step (b) utilizes a reverse transcription primer or a primer for DNA polymerization which ao corresponds is complementary to a contiguous sequence in the polynucleotide mold constituted by the single stranded target sequence and part of the nucleotide sequence 5 'or 3' adjacent.