福建农业学报
福建農業學報
복건농업학보
FUJIAN JOURNAL OF AGRICULTURAL SCIENCES
2014年
4期
345-349
,共5页
芒果%SRAP%体系优化%引物筛选%退火温度
芒果%SRAP%體繫優化%引物篩選%退火溫度
망과%SRAP%체계우화%인물사선%퇴화온도
Mango%SRAP%system optimization%primer selection%annealing temperature
以芒果为材料，通过单因子模板DNA含量、引物含量、2× Taq PCR Master Mix含量3种因素水平进行试验，建立了SRAP反应的优化体系。结果表明：最佳反应体系为20μL ，包括24 ng 模板DNA ，8 pmL引物，2× Taq PCR Master Mix 10.0μL。该体系对芒果扩增效果好，电泳结果显示扩增条带清晰，多态性高，而且操作简单。从81对引物组合中筛选出扩增条带清晰、多态性丰富的29对引物组合。在此基础上对退火温度进行探索，发现扩增结果对退火温度变化不敏感。
以芒果為材料，通過單因子模闆DNA含量、引物含量、2× Taq PCR Master Mix含量3種因素水平進行試驗，建立瞭SRAP反應的優化體繫。結果錶明：最佳反應體繫為20μL ，包括24 ng 模闆DNA ，8 pmL引物，2× Taq PCR Master Mix 10.0μL。該體繫對芒果擴增效果好，電泳結果顯示擴增條帶清晰，多態性高，而且操作簡單。從81對引物組閤中篩選齣擴增條帶清晰、多態性豐富的29對引物組閤。在此基礎上對退火溫度進行探索，髮現擴增結果對退火溫度變化不敏感。
이망과위재료，통과단인자모판DNA함량、인물함량、2× Taq PCR Master Mix함량3충인소수평진행시험，건립료SRAP반응적우화체계。결과표명：최가반응체계위20μL ，포괄24 ng 모판DNA ，8 pmL인물，2× Taq PCR Master Mix 10.0μL。해체계대망과확증효과호，전영결과현시확증조대청석，다태성고，이차조작간단。종81대인물조합중사선출확증조대청석、다태성봉부적29대인물조합。재차기출상대퇴화온도진행탐색，발현확증결과대퇴화온도변화불민감。
Mango seedlings were used as the test material in this experimentation to optimize SRAP by the single factor tests of primer DNA ,primer ,and 2xTaq PCR Master Mix . The results indicated that a 20 uL reaction system appeared to be most desirable ,as the resultant electrophoresis provided clear amplification bands and was highly polymorphic while the procedure was simple .From 81 primer combinations ,29 were selected with abundant polymorphism .The optimized SRAP-PCR system and polymorphism primer combinations could be applied for molecular genetic studies on mango .Our investigation showed that annealing temperature was not a significant factor on the amplification .
