生物技术通报
生物技術通報
생물기술통보
BIOTECHNOLOGY BULLETIN
2009年
12期
72-75
,共4页
林玲%汤浩茹%刘燕%候艳霞
林玲%湯浩茹%劉燕%候豔霞
림령%탕호여%류연%후염하
观赏桃%DNA提取%ISSR%体系优化
觀賞桃%DNA提取%ISSR%體繫優化
관상도%DNA제취%ISSR%체계우화
Ornamental peach%DNA extraction%ISSR%Optimal PCR system
采用改良CTAB法从观赏桃满天红叶片中提取基因组DNA,通过单因素实验探讨了模板DNA、Mg~(2+)、dNTPs和Taq DNA酶等条件对观赏桃ISSR-PCR扩增结果的影响,建立了ISSR-PCR扩增的最佳体系:2.5μl反应体系中包含10×Buffer 2.5μl,模板DNA 40ng,Mg~(2+)浓度2.5mmol/L,引物浓度04 μmol/L,dNTPs浓度0.4mmol/L,Taq DNA酶0.5U.利用所建立的体系对红叶桃、菊花桃和春艳等13份材料进行检验,其结果表明优化后的体系适合观赏桃的ISSR-PCR反应.
採用改良CTAB法從觀賞桃滿天紅葉片中提取基因組DNA,通過單因素實驗探討瞭模闆DNA、Mg~(2+)、dNTPs和Taq DNA酶等條件對觀賞桃ISSR-PCR擴增結果的影響,建立瞭ISSR-PCR擴增的最佳體繫:2.5μl反應體繫中包含10×Buffer 2.5μl,模闆DNA 40ng,Mg~(2+)濃度2.5mmol/L,引物濃度04 μmol/L,dNTPs濃度0.4mmol/L,Taq DNA酶0.5U.利用所建立的體繫對紅葉桃、菊花桃和春豔等13份材料進行檢驗,其結果錶明優化後的體繫適閤觀賞桃的ISSR-PCR反應.
채용개량CTAB법종관상도만천홍협편중제취기인조DNA,통과단인소실험탐토료모판DNA、Mg~(2+)、dNTPs화Taq DNA매등조건대관상도ISSR-PCR확증결과적영향,건립료ISSR-PCR확증적최가체계:2.5μl반응체계중포함10×Buffer 2.5μl,모판DNA 40ng,Mg~(2+)농도2.5mmol/L,인물농도04 μmol/L,dNTPs농도0.4mmol/L,Taq DNA매0.5U.이용소건립적체계대홍협도、국화도화춘염등13빈재료진행검험,기결과표명우화후적체계괄합관상도적ISSR-PCR반응.
The improved method CTAB was used to extract DNA from leaves of ornamental peach mantianhong.The research discussed different components effect to the DNA amplification,including program template,primer,dNTPs,Taq polymerase and Mg~(2+),through ladder experiments.The reaction conditions were optimized and then the optimal PCR system for ISSR analysis was established as follows,total volume 25 μl,25 μl 10 Buffer,40 ng template DNA,2.5 mmol/L Mg~(2+),0.4 mol/L Primer,0.4 mmol/L dNTPs,and 0.5 U Taq DNA polymerase.The optimal PCR system was tested by 13 samples,showing that it performed satisfactorily on ornamental peach.
