分子植物育种
分子植物育種
분자식물육충
MOLECULAR PLANT BREEDING
2013年
2期
262-266
,共5页
胡凤荣*%国荣荣%王斐%王江勇
鬍鳳榮*%國榮榮%王斐%王江勇
호봉영*%국영영%왕비%왕강용
东方百合%杂种后代%试管苗叶片%基因组DNA%提取方法
東方百閤%雜種後代%試管苗葉片%基因組DNA%提取方法
동방백합%잡충후대%시관묘협편%기인조DNA%제취방법
Oriental lily%Hybrid offspring%The leaves of in-tube seedling%Genomic DNA%Extraction method
以东方百合‘Sorbonne’בFrancia’的杂种后代试管苗叶片为材料,采用CTAB-高盐法、简易CTAB法、CTAB-硅珠法和SDS-CTAB法等4种DNA提取方法,比较DNA耗时、纯度、得率、浓度和质量等指标,以期为东方百合‘Sorbonne’בFrancia’杂种群体大量杂种后代试管苗叶片总DNA的提取筛选适宜方法.结果表明,除CTAB-硅珠法未获得DNA,其余3种方法均获得条带清晰、完整性好的DNA.在耗时方面, CTAB-硅珠法<简易CTAB法<CTAB-高盐法<SDS-CTAB法;在纯度方面,简易CTAB法纯度高于SDS-CTAB法、CTAB-高盐法,质量最佳,OD260/OD280为1.98,OD260/OD230为2.01,浓度为190.01 ng/μL,得率为57.01μg/g.综合比较,采用简易CTAB法进行杂种群体大量试管苗叶片总DNA提取效果更佳.
以東方百閤‘Sorbonne’בFrancia’的雜種後代試管苗葉片為材料,採用CTAB-高鹽法、簡易CTAB法、CTAB-硅珠法和SDS-CTAB法等4種DNA提取方法,比較DNA耗時、純度、得率、濃度和質量等指標,以期為東方百閤‘Sorbonne’בFrancia’雜種群體大量雜種後代試管苗葉片總DNA的提取篩選適宜方法.結果錶明,除CTAB-硅珠法未穫得DNA,其餘3種方法均穫得條帶清晰、完整性好的DNA.在耗時方麵, CTAB-硅珠法<簡易CTAB法<CTAB-高鹽法<SDS-CTAB法;在純度方麵,簡易CTAB法純度高于SDS-CTAB法、CTAB-高鹽法,質量最佳,OD260/OD280為1.98,OD260/OD230為2.01,濃度為190.01 ng/μL,得率為57.01μg/g.綜閤比較,採用簡易CTAB法進行雜種群體大量試管苗葉片總DNA提取效果更佳.
이동방백합‘Sorbonne’בFrancia’적잡충후대시관묘협편위재료,채용CTAB-고염법、간역CTAB법、CTAB-규주법화SDS-CTAB법등4충DNA제취방법,비교DNA모시、순도、득솔、농도화질량등지표,이기위동방백합‘Sorbonne’בFrancia’잡충군체대량잡충후대시관묘협편총DNA적제취사선괄의방법.결과표명,제CTAB-규주법미획득DNA,기여3충방법균획득조대청석、완정성호적DNA.재모시방면, CTAB-규주법<간역CTAB법<CTAB-고염법<SDS-CTAB법;재순도방면,간역CTAB법순도고우SDS-CTAB법、CTAB-고염법,질량최가,OD260/OD280위1.98,OD260/OD230위2.01,농도위190.01 ng/μL,득솔위57.01μg/g.종합비교,채용간역CTAB법진행잡충군체대량시관묘협편총DNA제취효과경가.
@@@@In order to screen the ideal method to extract genomic DNA from the Leaves of in-tube seedling of oriental lily, the hybrids offspring derived from the cross of Sorbonne × Francia were used as the experimental materials in this study. We compared the DAN quality and extracting efficiency of the employed DNA extraction methods, such as SDS-CTAB, Simple and ease CTAB, CTAB-high salt precipitation and CTAB-silica purifi-cation, in the facets of the purity, yield, concentration, and time consuming to choosing the ideal DNA extraction method. Three methods can be used to extract DNA with good banding profiles and Integrity except CTAB-silica purification missing DNA. The orders of the time consuming were from more to less following as CTAB-silica purification, Simple and ease CTAB, CTAB-high salt precipitation and SDS-CTAB. In the term of DNA purity, the simple CTAB method got the highest DNA purity than that of the SDS-CTAB and CTAB-high salt precipitation, which the extracted DNA with the excellent quality with OD260/OD280, 1.98, OD260/OD230, 2.01;while the concentration of the extracted DNA reached 190.01 ng/μL as well as 57.01μg/g of the DNA yield. Above all, Simple and ease CTAB is the most suitable way to extract a great quantity of genomic DNA from the leaves of in-tube seedling of the hybrid offspring of oriental lily.
