生物技术通报
生物技術通報
생물기술통보
BIOTECHNOLOGY BULLETIN
2010年
4期
112-115
,共4页
尤有利%王江波%施维属%潘东明
尤有利%王江波%施維屬%潘東明
우유리%왕강파%시유속%반동명
荔枝%RAPD%优化%PCR
荔枝%RAPD%優化%PCR
려지%RAPD%우화%PCR
Litchi chinensis%Sonn. RAPD%Optimize%PCR
为应用RAPD技术开展对荔枝种质资源的分析,以S43(GTCGCCGTCA)为引物,通过试验设计,分别研究了退火温度、模板浓度、引物浓度、dNTP浓度、Taq DNA聚合酶用量对荔枝RAPD-PCR反应的影响. 建立并优化了适宜荔枝RAPD分析的扩增体系:20 μL的反应体系,30 ng的模板DNA度,0.25 μmol/L RAPD引物、1.0 U Taq DNA聚合酶,0.2 μmol/L dNTP为荔枝适宜的RAPD-PCR扩增条件.
為應用RAPD技術開展對荔枝種質資源的分析,以S43(GTCGCCGTCA)為引物,通過試驗設計,分彆研究瞭退火溫度、模闆濃度、引物濃度、dNTP濃度、Taq DNA聚閤酶用量對荔枝RAPD-PCR反應的影響. 建立併優化瞭適宜荔枝RAPD分析的擴增體繫:20 μL的反應體繫,30 ng的模闆DNA度,0.25 μmol/L RAPD引物、1.0 U Taq DNA聚閤酶,0.2 μmol/L dNTP為荔枝適宜的RAPD-PCR擴增條件.
위응용RAPD기술개전대려지충질자원적분석,이S43(GTCGCCGTCA)위인물,통과시험설계,분별연구료퇴화온도、모판농도、인물농도、dNTP농도、Taq DNA취합매용량대려지RAPD-PCR반응적영향. 건립병우화료괄의려지RAPD분석적확증체계:20 μL적반응체계,30 ng적모판DNA도,0.25 μmol/L RAPD인물、1.0 U Taq DNA취합매,0.2 μmol/L dNTP위려지괄의적RAPD-PCR확증조건.
In order to use RAPD markers to analyze the resources in Litchi,the annealing temperature, template,concentration of primer, dNTP and the Taq DNA polymerase dosage on RAPD-PCR applications were tested to determine their optimal level . Besides,the following optimal reaction system for RAPD analysis in Litchi were established,including PCR reaction volume of 20 μL,30 ng DNA template,0.25 μmol/L RAPD primer,1.0 U Taq DNA polymerase,and 0.2 μmol/L dNTP in each reaction system of 20 μL.
