关键词: 甘蔗, 碱裂解, 中和, PCR模板, 多重PCR

Abstract: Using transgenic sugarcane leaves as materials, a small amount of young leaves were treated by alkali and transient heat, then neutralized, and cracking mixture was formed. The mixture was directly used as the template of PCR, and transgenic sugarcane exogenous gene of bar, KP4, CryIAc-2A-gna and endogenous gene Shactin as target genes, the amplified results were stable, accurate and reproducible, which reached the same effect of DNA amplification by conventional CTAB method. The templates by this method were still stable at room temperature for 2 weeks and at 4℃, -20℃ for 1 month. A series of exogenous gene PCR reactions were tested, which verified that this method was widely available in the detection of transgenic sugarcane. The method is fast for preparation of PCR templates without DNA extraction process, owing the advantages of requiring less material sample, low cost, simple, rapid, and efficient.

Key words: sugarcane, alkali lysis, neutralization, PCR template, multiple PCR