绿色科技
綠色科技
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LVSE DASHIJIU
2014年
3期
148-150
,共3页
墨兰%ISSR-PCR反应体系%优化
墨蘭%ISSR-PCR反應體繫%優化
묵란%ISSR-PCR반응체계%우화
Cymbidium sinense%ISSR-PCR reactionsystem%optimization
为建立墨兰的ISSR-PCR反应体系，采用正交试验分析了5个因素（Tag酶浓度、Mg2＋浓度、模板DNA浓度、引物浓度、dNTP浓度）在4个水平上对 ISSR -PCR反应的影响。结果表明：Tag 酶浓度为1.6U/25μL、Mg2＋浓度为2.4mmol/L、dNTP浓度0.96 mmol/L、Primer浓度为0.64μmol/L、DNA 浓度3.2ng/25μL是墨兰ISSR-PCR最优反应体系。
為建立墨蘭的ISSR-PCR反應體繫，採用正交試驗分析瞭5箇因素（Tag酶濃度、Mg2＋濃度、模闆DNA濃度、引物濃度、dNTP濃度）在4箇水平上對 ISSR -PCR反應的影響。結果錶明：Tag 酶濃度為1.6U/25μL、Mg2＋濃度為2.4mmol/L、dNTP濃度0.96 mmol/L、Primer濃度為0.64μmol/L、DNA 濃度3.2ng/25μL是墨蘭ISSR-PCR最優反應體繫。
위건립묵란적ISSR-PCR반응체계，채용정교시험분석료5개인소（Tag매농도、Mg2＋농도、모판DNA농도、인물농도、dNTP농도）재4개수평상대 ISSR -PCR반응적영향。결과표명：Tag 매농도위1.6U/25μL、Mg2＋농도위2.4mmol/L、dNTP농도0.96 mmol/L、Primer농도위0.64μmol/L、DNA 농도3.2ng/25μL시묵란ISSR-PCR최우반응체계。
In order to establish theISSR -PCR reaction systemofISSR -PCR reactionsystem ,this article use-sorthogonalexperiment to analyze the influences of fivefactors including Tagenzyme concentration ,Mg2+ con-centration ,DNA templateconcentration ,primer concentration ,and dNTP concentrationatfourlevelsof the or-thogonal teston theCymbidiumISSR -PCRreaction system .The results show thatwhen Tag enzyme concen-trationis 1 .6U/25μl ,M g2+ concentration is 2 .4mmol/L ,dN T Pconcentration is 0 .96 mmol/L ,concentration of primeris 0 .64μmol/L ,DNAconcentrationis 80ng/25μl ,theISSR-PCRreaction system is optimal .
