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时间:2020-05-20
《适合MSAP分析的核桃子叶DNA提取方法-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、林业科学研究2014,27(1):128~132ForestResearch文章编号:1001-1498(2014)01-0128-05适合MSAP分析的核桃子叶DNA提取方法周贝贝,陈凌娜,张俊佩,裴东(中国林业科学研究院林业研究所,林木遗传育种国家重点实验室,北京100091)关键词:核桃;子叶;DNA提取;多糖;MSAP中图分类号:$718.46文献标识码:ADNAExtractionMethodofWalnutCotyledonSuitableforMSAPAnalysisZHOUBei—bei,CHENLing—M,ZHANGJun-pei,PElDon
2、g(ResearchInstituteofForestry,ChineseAcademyofForestry,StateKeyLaboratoryofTreeGeneticsandBreedingofStateForestryAdministration,Beijing100091,China)Abstract:Toobtainhigh-qualityDNAsuitableformethylation-sensitiveamplificationpolymorphism(MSAP),a—nalysis,thematurecotyledonandtissuecult
3、uredcotyledonofwalnutcuhivar‘Liaoning一1’wereusedasthetestingmaterialsintheresearchofDNAisolationby3methods(SDS,highsaltCTABandmodifiedhighsaltCTAB)andthematureleartissuewastakenasthecontro1.TheconcentrationandintactnessoftheextractedDNAweredeter.minedusingaspectrometerandbyelectrophor
4、esisin0.8%(w/v)agarosegelsagainststandardsolutionsoflambdaDNA.HighqualityDNAwasextractedfromleaftissuesusingthehighsaltCTABmethodandthemodifiedhighsaltCTABmethod.NodetectableDNAwasextractedfromthematurecotyledonandcuhuredcotyledonusingSDSmeth—od.TheDNAwithhighqualitybutlittlequantityw
5、asextractedfromthematurecotyledonbutnottheextractcul—turedcotyledonusingthehighsaltCTABmethod.TheDNAwithhighquantityandqualitysuitableforMSAPanaly—sisfrombothmaturecotyledonandculturedcotyledonwasextractedusingthemodifiedhighsaltCTABmethod.TheDNAextractionmethodofwalnutcotyledonsuitab
6、leforMSAPanalysiswasestablished,whichcouldlayfoundationfordownstreammolecularbiologyexperimentandprovideareferencetothepolysaccharideremovalresearchinDNAextractionprocedure.Keywords:Juglansregia;cotyledon;DNAextraction;polysaceharide;MSAP核桃(JuglansregiaL.)营养价值高,含有丰富后续实验造成较大影响J。目前,核桃组织
7、全基因的脂肪酸和蛋白质J,子叶是核桃重要的营养储存组DNA提取方法主要集中在叶片组织。雷玲器官,作为常用的实验材料,可用来组织培养进行生等_6以13个河北核桃优系叶片为材料,比较了高盐根实验等相关研究-3]。MSAP是在AFLP基础上低pH值法、SDS法和CTAB法提取基因组DNA的建立起来的在全基因组范围检测5’一CCGG位点的效果,确定含3%PVP的高盐低pH值法为河北核桃胞嘧啶甲基化变化的实验技术J,具有多态性高、无叶片DNA提取的最适方法;然而,核桃子叶的DNA需预知DNA序列的优点,但其需要提取高质量的基提取目前没有较为有效的方法。本试验以‘辽宁1因组D
8、NA,存在
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