Establishment of AFLP Analysis System in Spinach (Spinacia oleracea L.)

In this study total genetic DNA from tender leaves of spinach (Spinacia oleracea) was extracted with improved CTAB. An AFLP (amplified fragment length polymorphism) analysis system was established by optimizing the concentration of template DNA, DNA restriction reaction, ligase reaction, pre-amplification and selective amplification. The results were obtained as follow: (1) The effect of restriction enzyme was better in PCR instrument than in water-bath, the reaction time and the concentration of DNA were not sensitive to restriction enzyme. (2) The best reaction system in 20 mu l total volume contained Mg2+ (25 mmol/L) 1.2 mu l, dNTPs (2.5 mmol/L) 1.6 mu l and Taq-polymerase (5U/mu l) 0.2 mu l. This paper provided a method for further study in the field of genetic relative analysis of spinach (Spinacia oleracea L.) cultivars and genetic breeding.