贵州农业科学
貴州農業科學
귀주농업과학
GUIZHOU AGRICULTURAL SCIENCES
2015年
2期
29-34
,共6页
王惠君%王文泉%卢诚%王海燕%陈新
王惠君%王文泉%盧誠%王海燕%陳新
왕혜군%왕문천%로성%왕해연%진신
橡胶树%SRAP%体系优化%引物组合筛选
橡膠樹%SRAP%體繫優化%引物組閤篩選
상효수%SRAP%체계우화%인물조합사선
Hevea brasiliensis%SRAP%system optimization%primer combinations screening
为橡胶树遗传多样性分析及遗传图谱的构建奠定基础,分析引物浓度、Mg2+浓度、dNTPs 浓度、模板 DNA 用量、Taq 酶用量等因素对 SRAP-PCR 扩增反应的影响。结果表明,橡胶树 SRAP-PCR 的适宜扩增反应体系(20μL)为:引物0.2μmol/L,Mg2+2.0 mmol/L,dNTPs 0.20 mmol/L,模板 DNA50 ng, Taq 酶0.5 U。利用该体系从400对引物组合中初筛出28组多态性好的引物组合,获得250条清晰带,多态性比率为62.8%。
為橡膠樹遺傳多樣性分析及遺傳圖譜的構建奠定基礎,分析引物濃度、Mg2+濃度、dNTPs 濃度、模闆 DNA 用量、Taq 酶用量等因素對 SRAP-PCR 擴增反應的影響。結果錶明,橡膠樹 SRAP-PCR 的適宜擴增反應體繫(20μL)為:引物0.2μmol/L,Mg2+2.0 mmol/L,dNTPs 0.20 mmol/L,模闆 DNA50 ng, Taq 酶0.5 U。利用該體繫從400對引物組閤中初篩齣28組多態性好的引物組閤,穫得250條清晰帶,多態性比率為62.8%。
위상효수유전다양성분석급유전도보적구건전정기출,분석인물농도、Mg2+농도、dNTPs 농도、모판 DNA 용량、Taq 매용량등인소대 SRAP-PCR 확증반응적영향。결과표명,상효수 SRAP-PCR 적괄의확증반응체계(20μL)위:인물0.2μmol/L,Mg2+2.0 mmol/L,dNTPs 0.20 mmol/L,모판 DNA50 ng, Taq 매0.5 U。이용해체계종400대인물조합중초사출28조다태성호적인물조합,획득250조청석대,다태성비솔위62.8%。
To lay the foundation for evaluation of genetic diversity analysis and genetic linkage map in H.brasiliensis,factors influencing SRAP-PCR amplification system were analyzed,including primer concentration,Mg2+ concentration,dNTPs concentration,DNA template dosage and Taq polymerase. Results:A suitable SRAP-PCR analysis reaction system for H.brasiliensis was obtained.The reaction mixture (20 μL)contained primer 0.2 μmol/L,Mg2+ 2.0 mmol/L,dNTPs 0.20 mmol/L,50 ng of genomic DNA template,0.5 U of Taq polymerase.Conclusion:using optimized SRAP-PCR system,28 pairs of primers were screened out of 400 pairs of primers. 250 clear bands were gained and the polymorphism rate was 62.8%.