The invention discloses a
cordyceps militaris fruiting body development stage internal
reference gene, a primer, a
screening method and application, wherein the internal
reference gene is TUB, and thenucleotide sequence is SEQ ID NO: 1. The
nucleotide sequences of the forward and reverse primers of TUB are SEQ ID NO: 13-14. The
screening method comprises the following steps: (1) selecting twelveinternal
reference genes as candidate internal
reference genes, and designing primers; (2) inducing
cordyceps militaris fruiting body development, extracting total RNAs at different development stages, measuring
RNA concentration, carrying out reverse transcription on the
RNA to synthesize cDNA, taking the cDNA as a template, and analyzing the candidate internal
reference genes by using real-timefluorescence quantitative PCR; and (3) carrying out Ct value analysis on the real-time
fluorescence quantitative PCR data to screen out stable internal reference genes. According to the application, the TUB is used as the internal
reference gene to detect the relative expression levels of target genes at different development stages. The stability of the internal reference
gene is good; and the method is simple, rapid and low in cost.