SUN Yong-jian,SUN Ning,CAO Zhi-pan,et al.Study on DNA Extraction of Saintpaulia ionantha for AFLP Reaction System[J].Northern Horticulture,2012,36(22):104-107.
适于AFLP分析的非洲紫罗兰DNA提取方法研究
- Title:
- Study on DNA Extraction of Saintpaulia ionantha for AFLP Reaction System
- 文章编号:
- 1001-0009（2012）22-0104-04
- Keywords:
- DNA extraction; Saintpaulia ionantha; CTAB; freeze-drying; AFLP analysis
- 分类号:
- S 681.2
- 文献标志码:
- A
- 摘要:
- 以非洲紫罗兰同一植株的叶片和茎段为试材，采用新鲜组织提取、烘干组织提取、冷冻干燥组织提取的3种不同预处理方法，并采用DNA Kit、CTAB-Ⅰ、CTAB-Ⅱ 3种提取方法，比较分析了DNA片断大小及不同方法提取DNA的纯度与得率，以期确定适用于多酚、多糖、水分含量高的非洲紫罗兰的DNA提取方法。结果表明：采用冷冻干燥组织提取法，经改良的CTAB-Ⅱ法，即在2次抽提步骤中，加入一步利用CTAB沉淀液沉淀DNA的方法最适合非洲紫罗兰DNA提取。这样得到的基因组DNA可作为后续分子水平研究的优良PCR模板。
- Abstract:
- Taking the same leaves and stems of Saintpaulia ionantha as materials,fresh tissue extraction,drying tissue extraction,freeze drying tissue extraction,3 different pre-treatment were adapted,also DNA Kit,CTAB-Ⅰ,CTAB-Ⅱ 3 different extraction methods were adopt,to compare DNA fragment size,purity and yield throagh different extraction method.The best DNA extraction method for Saintpaulia ionantha which contained more polysaccharide polyphenols and moisture was determined.The results showed CTABⅡ method and the material processed by freeze-drying before test,namely,in the second extraction steps,added CTAB preeipitation in the method of DNA precipitation methed.DNA received by this method could be used as template for PCR in study at further molecular research.
参考文献/References:
[1]吴忠兰,宋玉霞,马洪爱,等.用改进的CTAB法提取肉苁蓉基因组DNA[J].宁夏大学学报(自然科学版),2008,29(1):78-80. [2]Porebski S，Baileyl G，Bernard R.Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components[J].Plant Molecular Biology Reporter,1997,15(1):8-15. [3]谢让金,邓烈.一种适合AFLP分析的柑橘DNA提取方法[J].生物技术,2007,17(6):27-28. [4]Pierre G，Haurencem D.Isolation of plant DNA：a fast，inexpensive，and reliable method[J].Plant Mol Biol Rep,1992,10:60-65. [5]熊光明,梁国鲁,阎勇,等.适用于AFLP分析用的柑橘DNA提取方法[J].果树学报,2002,19(4):267-268. [6]许庆霖.非洲紫罗兰-最可人的室内花卉（上）[J].中国花卉盆景,1998(11):22-23. [7]林子.非洲紫罗兰的温室规模化生产[J].中国花卉园艺,2007(2):10-13. [8]孙永健.非洲紫罗兰核型及DNA甲基化表观遗传作用的研究[D].天津:天津农学院,2010. [9]李海渤.茴香总DNA提取方法的比较研究[J].北方园艺,2008(3):189-192. [10]黄永芳,廖国春,吴雪辉,等.一种适合AFLP分析的油茶DNA提取方法[J].广东林业科技,2009,25(1):23-26. [11]许婉芳.去除顽拗植物DNA提取过程中干扰物质的方法[J].闽西职业大学学报,2002(3):56-58. [12]程志学,陈清华,于远,等.适合AFLP分析用的节瓜基因组DNA提取方法的研究[J].分子植物育种,2009,7(2):420-424.
相似文献/References:
[1]陈 娟,金文龙,莫亿伟.两种方法提取板栗叶片DNA的效果比较[J].北方园艺,2013,37(18):89.
CHEN Juan,JIN Wen-long,MO Yi-wei.Comparison on Genomic DNA Extracted from the Leaves of Chinese Chestnut by Two Methods[J].Northern Horticulture,2013,37(22):89.
[2]刘晓东,王婷婷,刘群录,等.抗生素对非洲紫罗兰不定芽再生的影响[J].北方园艺,2014,38(03):80.
LIU Xiao-dong,WANG Ting-ting,LIU Qun-lu,et al.Effect of Antibiotics on Adventitious Shoot Regeneration of Saintpaulia ionantha [J].Northern Horticulture,2014,38(22):80.
[3]赵志常,陈业渊,高爱平,等.改良CTAB法提取番石榴总DNA的初步研究[J].北方园艺,2013,37(09):123.
ZHAO Zhi-chang,CHEN Ye-yuan,GAO Ai-ping,et al.Preliminary Study on the Total DNA Extracted from Psidium guajava L. by Improved CTAB Method[J].Northern Horticulture,2013,37(22):123.
[4]陈名红,李 玉,刘 多,等.适于ISSR-PCR扩增的百合基因组DNA的提取[J].北方园艺,2012,36(24):108.
CHEN Ming-hong,LI Yu,LIU Duo,et al.Extraction of Genomic DNA for ISSR-PCR Reaction in Lilium[J].Northern Horticulture,2012,36(22):108.
[5]裴 仁 济,陈 小 强,孙 宁,等.花色模式植物—非洲紫罗兰[J].北方园艺,2010,34(11):0.[doi:10.11937/bfyy.2010011078]
PEI Ren -ji,CHEN Xiao -qiang,SUN Ning,et al.Flower Colors Model Plant-Saintpqulia ionantha[J].Northern Horticulture,2010,34(22):0.[doi:10.11937/bfyy.2010011078]
[6]李 志 虹,高 英,倪 苏,等.秋水仙素离体诱导对非洲紫罗兰形态的影响[J].北方园艺,2010,34(05):0.[doi:10.11937/bfyy.201005057]
LI Zhi -hong,GAO Ying,NI Su,et al.The Effect of Colchicine on the Formof Saintpauliaionantha in vitro[J].Northern Horticulture,2010,34(22):0.[doi:10.11937/bfyy.201005057]
[7]陈芸,高原青,王继莲,等.不同保存方法对石榴成熟叶片基因组DNA提取效果的影响[J].北方园艺,2016,40(08):100.[doi:10.11937/bfyy.201608028]
CHEN Yun,GAO Yuanqing,WANG Jilian,et al.Effect of Different Preservation Methods on Genomic DNA Extraction of Pomegranate[J].Northern Horticulture,2016,40(22):100.[doi:10.11937/bfyy.201608028]
[8]徐 宏 英,陈 会 仙,岳 建 英,等.非洲紫罗兰有机型无土栽培研究[J].北方园艺,2004,28(04):0.[doi:10.11937/bfyy.200404062]
[J].Northern Horticulture,2004,28(22):0.[doi:10.11937/bfyy.200404062]
[9]何家涛,王 会.非洲紫罗兰幼嫩叶片离体培养的技术研究[J].北方园艺,2007,31(01):0.[doi:10.11937/bfyy.200701071]
[10]刘伟,赵琬玥,周长梅,等.绿绒蒿属植物不同DNA提取方式的比较[J].北方园艺,2017,41(15):97.[doi:10.11937/bfyy.20164449]
LIU Wei,ZHAO Wanyue,ZHOU Changmei,et al.Camparison of Different DNA Extraction Methods for Detection of Meconpsis Vig[J].Northern Horticulture,2017,41(22):97.[doi:10.11937/bfyy.20164449]
备注/Memo
第一作者简介：孙永健（1983-），男，硕士，助教，现主要从事生物化学及分子生物学方面的教学与研究工作。E-mail：syjwonderful@yahoo.com.cn.