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《Bulletin of The Academy of Military Medical Sciences》 2001-03
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Optimal scheme of reaction conditions for randomly amplified po lymorphic DNA of Pseudomonas aeruginosa

YE MingLiang1, WANG QuanLi2(1.Jinan Military General Hospital, Jinan 250031, China; 2.Institute of Field Transfusion, Academy of Military Medical Sciences, Beijing 100850, Chin a)  
ObjectiveTo obtain optimum scheme of reaction c on ditions for randomly amplified polymorphic DNA(RAPD) of Pseudomonas aer uginosa~{!~}WT~{!=~}. MethodsThe optimal reaction systems of RAPD wer e obtained by single factor design and response surface design. The effects of d ifferent types of thermocycler,and temperatures of annealling and extension on RAPD profile were compared. Results:~{!~}WTB Z~{!=~} Multiple optimal reaction systems caused interaction of multifactors were ob served in single factor design and the concentrations of key components of the o ptimal reaction system with relative stability obtained by response surface desi gn were ~{*)~}2.0?mmol/L~{**~} of Mg~{*,*)~}2+~{**~}, 0.5?~{L~}mol/L of primer and 0.5?ng/~{L~}l of template. The optimal temperatures of annealling and extension were 38-43~{!f~} and 62-72~{!f~} respectively. Conclusions : The best scheme to optimize the reaction system of RAPD was response s urface d esign. The optimal temperatures of annealling and extension in amplification con ditions were 38-43~{!f~} and 62-72~{!f~} respectively.
【Fund】: 全军医药卫生“九五”青年基金资助课题( 98Q0 12 )
【CateGory Index】: R378
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