Optimization and Establishment of AFLP Analying System in Spinach(Spinacia oleracea L.)

doi:10.7668/hbnxb.2010.04.024

Abstract

Abstract: In this study toal 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 follows: ①The effect of restriction enzyme was better in PCR instrument than in waterbath， a nd the reaction time and the concentration of DNA were not sensitive to restriction enzyme. ②The best reaction system in 20 μL total volume contained Mg²⁺ (25 mmol /L) 1. 2 μL，dNTPs (2. 5 mmol /L) 1. 6 μL and Taq-polymerase(5 U /μL) 0. 2 μL. This paper provided a method for further study in the field of genetic relative analysis of Spinach (Spinacia oleracea L.) varieties and genetic breeding.

Key words: Spinach (Spinacia oleracea L.), AFLP, Influencing factors

CLC Number:

S636.1

MEI Yi, HOU Lei-ping, CUI Yan-ling, CHEN Hai-li, MENG Shu-chun. Optimization and Establishment of AFLP Analying System in Spinach(Spinacia oleracea L.)[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(4): 111-115. doi: 10.7668/hbnxb.2010.04.024.

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