Total DNA Extraction and RAPD System Optimization of Tamarix Plants in Tarim Basin Desert
In this research, the extraction method for total DNA from young leaves of Tamarix plants growing in arid, desert, saline-alkali habitat was expored and optimized their reaction system and amplification conditions of RAPD-PCR.The results showed that the total DNA extracted had good quality by using the improved method based on CTAB-high salt precipitation and its yield were also high significantly, so the method could be fit for the total DNA extraction of most plants in Tamarix L..At the same time, by one-factor comparative experiments, the suitable RAPD-PCR system for Tamarix plants was developed as follows: in 25 μL PCR solution, including 2.5 mmol/L Mg2+, 0.25 mmol/L dNTPs, 0.30 μmol/L primer, 1.5 U Taq DNA polymerase, 200 ng DNA template.Moreover, the amplification conditions were 95℃ for 4 min, 40 cycles of 94℃ for 30 s, 36℃ for 40 s, and 72℃ for 1 min and a terminal extension step at 72℃ for 10 min, and then kept at 4℃.In addition, through the confirmatory test with six polymorphic primer pairs, the optimized reaction system could amplify stably and its amplification conditions can meet the requirement for molecular polymorphic analysis of most Tamarix L.plants.
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NSTL
万方数据
维普
