Establishment and optimization of AFLP reaction system in Asarum sieboldii / 中草药

ABSTRACT

Objective To establish and optimize AFLP reaction system used in providing necessary technique basis for genetic diversity analysis in Asarum sieboldii.Methods Leaves of A.sieboldii were used as experimental materials to analyze various essential elements of the whole process,such as quality of extracted DNA,time of restriction digest and ligation,concentration of Mg2+,primers and Taq DNA polymerase during PCR amplification etc.,so that the most optimal AFLP reaction system could be built up. Results The optimal AFLP reaction system of A.sieboldii has been constructed in the genomic DNA extraction,mercaptoethanol being utilized and samples being incubated about 30 min at 65 ℃;the genomic DNA being digested by Trul Ⅰ and Pst Ⅰ for 3 h respectively;in PCR amplification,the final concentration of Mg2+ being 1.5 mmol/L and the volume of Taq DNA polymerase being 0.2 ?L.Conclusion The present reaction system for A.sieboldii is able to gain the favorable results of AFLP analysis and can be used in the genetic diversity research of the species.