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1.
利用改良的CTAB方法从香蕉叶片中提取高质量的DNA。在参考一般RAPD分析反应程序的基础上,经过反复试验,确定适合香蕉PCR扩增程序为:94℃预变性 5min;94℃变性 1min;38℃退火 1min;72℃延伸 2min;变性ó延伸,循环45次;最后72℃延伸 2min。PCR扩增的体系(总体积25μL)为:模板DNA 20ng,dNTP 200μmol/L,10×PCR Buffer2.5μL,引物0.20μmol/L,Taq酶0.75U,ddH2O17.85μL。  相似文献   

2.
以鼠尾草属植物叶片为材料,提取基因组DNA,并对RAPD反应条件进行了系统优化.结果表明,采用核DNA法提取的DNA质量较高,适宜于RAPD分析;RAPD扩增最佳反应体系为20μl反应体系中,10×buffer 2.0μl,模板DNA 20 ng,Mg 浓度2.0 mmol/L,引物浓度0.6μmol/L,dNTPs浓度0.2mmol/L,Taq酶1.0U.扩增反应程序为94℃预变性5min,94℃变性1min,36℃退火1main,72℃延伸2main;40个循环;72℃后延伸10main,4℃保存.  相似文献   

3.
长筒石蒜花被片DNA的提取及ISSR体系的建立   总被引:9,自引:0,他引:9  
以长筒石蒜为例,探讨了花被片DNA的提取方法。并在此基础上,确立其ISSR反应程序及体系。反应程序为: 94℃预变性3min,进入38个PCR循环( 94℃变性30s, 58℃复性30s, 72℃延伸90s),最后于72℃延伸7min。扩增反应总体积为20μL: 1×Taq酶扩增缓冲液, 1. 5mmol/LMg2+, 200μmol/LdNTP, 0. 5μmol/L随机引物, 0. 5UTaq聚合酶,DNA模板10~30ng。  相似文献   

4.
以桃基因组DNA为模板,通过正交试验设计,从Mg2+、Taq酶、dNTP、引物、模板5种因素4个水平对桃SRAP反应体系进行优化,建立了适合于桃的SRAP-PCR优化反应体系,该25μL反应体系:模板DNA50 ng,MgCl22.5 mmol/L,dNTP200μmol/L,上下引物各0.4μmol/L,Taq DNA聚合酶1.5 U,以灭菌双蒸水补齐至25μL。PCR反应程序为:94℃预变性5min;94℃变性1 min,35℃复性1 min,72℃延伸1 min,5个循环;94℃变性1 min,50℃复性1 min,72℃延伸1 min,35个循环,72℃延伸10 min。  相似文献   

5.
荸荠基因组DNA的提取及RAPD反应体系的建立   总被引:4,自引:1,他引:3  
以荸荠叶状茎为试验材料,采用改良的SDS法提取其基因组DNA,并对其RAPD反应体系进行优化,建立了荸荠的RAPD—PCR优化反应体系和程序。结果表明,提取的基因组DNA纯度和完整性较好,OD260/OD280值在1.8~2.0之间,DNA无降解现象,完全可以满足RAPD—PCR扩增要求。建立了荸荠RAPD反应体系:总体积为25μl,各有关成分的最佳浓度分别为25mmol/L Mg^2+,1.0UTaqDNA聚合酶,0.2mmol/L dNTPs,1μmol/μl引物,1.5ng/μl DNA模板。PCR反应程序为:94%预变性3min;94℃变性1min,37℃退火30s,72℃延伸60s,40个循环;最后72%延伸10min。  相似文献   

6.
为确保樱花RAPD扩增结果的稳定性和重复性,对Mg2 、dNTP、引物、模板DNA浓度、Taq酶用量、退火温度,以及PCR循环次数等影响樱花RAPD结果的重要因素进行了初步探索。试验表明,樱花RAPD扩增最适反应体系为20μl反应液中:1×PCR buffer,2.5 mmol/L MgCl2,0.15 mmol/L dNTP,1 UTaq酶,0.2μmol/L 10bp随机引物,20~30 ng模板DNA。最佳扩增程序为:94℃预变性4 min,94℃变性30 s,38℃退火30 s,72℃延伸2 min,循环40次,最后72℃延伸10 min,12℃保存。  相似文献   

7.
百合鳞片DNA提取及RGA-PCR体系的优化   总被引:4,自引:0,他引:4  
以百合鳞片为材料,采用改良的CTAB法获得了高质量的基因组DNA.通过影响PCR反应各因子的优化,建立了适合百合的RGA-PCR反应体系:25μl体系中含30 ng模板DNA、2.5 mmol/L MgCl2、0.2 mmol/L d.NTPs、0.6 μmol/L引物及1.5 U Taq酶.扩增程序为:94 ℃预变性5 min,94℃变性1 min,44 ℃退火1 min,72 ℃延伸2 min,40个循环;最后72 ℃延伸7 min.利用该体系对35个百合品种进行RGA-PCR,表明该反应体系具有较好的稳定性和可靠性.  相似文献   

8.
春兰ISSR-PCR反应体系的优化   总被引:11,自引:0,他引:11  
以春兰为材料,对影响ISSR-PCR扩增结果的因素如模板DNA、dNTPs、MgCl2、primer、TaqDNA聚合酶的浓度及引物退火温度、反应循环数进行了探讨,确立了适合春兰ISSR-PCR分析的最佳反应体系和PCR扩增参数:在15μL反应体系中含1×buffer,0.2 mmol/L dNTPs,2.0 mmol/L MgCl2,0.2μmol/L引物,0.3 UTaqDNA聚合酶,20 ng模板DNA。PCR扩增程序:94℃预变性2.5 min;94℃变性45 s,48~58℃退火(退火温度随引物不同而定)45 s,72℃延伸2 min,35个循环;72℃延伸5 min。  相似文献   

9.
人参黑斑病菌RAPD反应体系的优化   总被引:1,自引:0,他引:1  
采用改良的CTAB法提取人参黑斑病菌的基因组DNA,建立人参黑斑病菌RAPD反应的体系.最佳体系容积为25μL,其中包括10×Taq配套缓冲液2.5μL,模板DNA 20 ng/μL,引物15 pmol/L,dNTP 150μmol/L,Taq DNA聚合酶1 U,Mg2+1.5 mmol/L,其余部分用DW补充.PCR扩增程序为:94℃预变性5 min,94℃变性1 min,40℃退火1 min,72℃延伸2 min,40次循环,72℃延伸7 min.  相似文献   

10.
正交设计优化番茄基因组DNA SSR-PCR反应体系   总被引:2,自引:0,他引:2  
以21份番茄品种为材料,应用正交试验设计对影响番茄SSR—PCR的主要参数进行优化,建立适于番茄的SSR反应体系和扩增程序。在15μl体系中各反应的最适合含量为:2.5μl ddH2O,1μl 40ng模板DNA,3μl 200μmol/L dNTP,3μl 0.8μmol/L SSR引物,0.1μl 0.5U/μl Taq DNA聚合酶,1.5μl 10×PCR Buffer,0.9μl 1.5mmol/L MgCl2。PCR适宜扩增程序为:94℃预变性2min;94℃变性1min,45℃退火25s,共35个循环;68℃延伸25s,68℃延伸10min,然后4℃保存。利用此反应体系对21个番茄品种进行PCR扩增并电泳检测,扩增结果清晰且有较高的多态性,表明该体系适合番茄的遗传多样性研究。  相似文献   

11.
分析了现有龙眼冻害分级标准中存在的问题,以及制订龙眼冻害分级标准应考虑的因素,在此基础上制订新的龙眼冻害分级标准,并提出了各冻害级别应采取的生产技术措施.  相似文献   

12.
综述氯酸钾对龙眼催花的效应及其机理,概述应用氯酸钾促使龙眼开花应注意及存在的问题。  相似文献   

13.
[目的]优化龙眼AFLP银染体系中的影响因素。[方法]以23份龙眼品种和2份龙眼近缘植物龙荔为试材,通过对扩增片段长度多态性(AFLP)反应体系中的几个影响因素(DNA提取、酶切与连接、电泳与银染)进行研究,建立起龙眼AFLP银染分析体系。[结果]用紫外分光光度计检测样品DNA质量和浓度,发现提取的DNA吸光度A260/A280比值在1.8左右,A260/A230的比值大于2.0,说明所提取的DNA样品纯度较高,无蛋白质或盐类的污染。酶切与连接中采用37℃水浴5 h,然后转入65℃水浴2 h的方法能保证酶切充分,又能防止酶切过度。电泳和银染时用4℃预冷的显影液进行显影,可以降低显影速度。[结论]采用该技术体系得到的AFLP指纹式样清晰,重复性强,适宜用作龙眼品种鉴别和遗传多样性分析。  相似文献   

14.
特殊气候环境下龙眼高产栽培技术   总被引:2,自引:0,他引:2  
经多年种植龙眼的技术服务与实践,探索出一套适宜边缘地带龙眼高产优质的栽培技术措施:科学施肥,满足果树生长需要;适时放梢、培育健壮晚秋梢作结果母枝;做好树体管理;控冬梢促花;保花保果,提高座果率;防治病虫,保护果树;保树体及防冻害技术措施。  相似文献   

15.
[Objective] The study aimed to provide supports for developing chilling and freezing injury monitoring and disaster damage assessment of longan(Dimocarpus Longan Lour.).[Method] Based on field observation data,the relationships between longan canopy temperature and air temperature under different weather types(sunny,cloudy to sunny,cloudy,rainy,radiation chilling injury and advection chilling injury)in 2007-2008 winter were analyzed.[Result] Diurnal variations of longan canopy temperature under sunny and radiation chilling injury weather conditions were most dramatic,followed with those under cloudy to sunny condition,while variations under cloudy,rainy and advection chilling injury conditions were mild.Diurnal variations of orchard air temperature were also closely related to weather types.By using linear and curvilinear regression methods,the relationship models between longan canopy temperature and observation station air temperature were established.The models for cloudy,rainy and advection chilling injury had better effects than those for sunny,cloudy to sunny and radiation chilling injury;the models for night were better than those for daytime and the whole day.[Conclusion] To some extent,applying the relationship models between longan canopy temperature and observation station air temperature could make up the shortcoming of meteorological data which were higher than the real values.  相似文献   

16.
龙眼越冬期间冠层温度与大气温度关系的初步分析(英文)   总被引:2,自引:0,他引:2  
[Objective] The study aimed to provide supports for developing chilling and freezing injury monitoring and disaster damage assessment of longan(Dimocarpus Longan Lour.).[Method] Based on field observation data,the relationships between longan canopy temperature and air temperature under different weather types(sunny,cloudy to sunny,cloudy,rainy,radiation chilling injury and advection chilling injury)in 2007-2008 winter were analyzed.[Result] Diurnal variations of longan canopy temperature under sunny and radiation chilling injury weather conditions were most dramatic,followed with those under cloudy to sunny condition,while variations under cloudy,rainy and advection chilling injury conditions were mild.Diurnal variations of orchard air temperature were also closely related to weather types.By using linear and curvilinear regression methods,the relationship models between longan canopy temperature and observation station air temperature were established.The models for cloudy,rainy and advection chilling injury had better effects than those for sunny,cloudy to sunny and radiation chilling injury;the models for night were better than those for daytime and the whole day.[Conclusion] To some extent,applying the relationship models between longan canopy temperature and observation station air temperature could make up the shortcoming of meteorological data which were higher than the real values.  相似文献   

17.
[Objective] The study aimed to provide supports for developing chilling and freezing injury monitoring and disaster damage assessment of longan(Dimocarpus Longan Lour.).[Method] Based on field observation data,the relationships between longan canopy temperature and air temperature under different weather types(sunny,cloudy to sunny,cloudy,rainy,radiation chilling injury and advection chilling injury)in 2007-2008 winter were analyzed.[Result] Diurnal variations of longan canopy temperature under sunny and radiation chilling injury weather conditions were most dramatic,followed with those under cloudy to sunny condition,while variations under cloudy,rainy and advection chilling injury conditions were mild.Diurnal variations of orchard air temperature were also closely related to weather types.By using linear and curvilinear regression methods,the relationship models between longan canopy temperature and observation station air temperature were established.The models for cloudy,rainy and advection chilling injury had better effects than those for sunny,cloudy to sunny and radiation chilling injury;the models for night were better than those for daytime and the whole day.[Conclusion] To some extent,applying the relationship models between longan canopy temperature and observation station air temperature could make up the shortcoming of meteorological data which were higher than the real values.  相似文献   

18.
从东璧龙眼优良单株取接穗,采用高接换种方法对福眼龙眼进行品种改良。从园地选择、土壤管理、科学施肥、疏花疏果、整形修剪、控冬梢促花、保花保果、病虫害防治等技术环节,总结其优质高效栽培技术。  相似文献   

19.
针对当前我国龙眼鲜果价格低迷、产量不稳、销售渠道不畅、效益滑坡的现状,指出需对龙眼生长结实特性、经营品种、经营投入与产出、栽培管理关键技术等知识重新认识,提出成功经营龙眼园要充分利用土地、水资源、气候资源以及资金、劳力、技术等资源要素,综合运筹时间价格差异、产品质量、果品加工和诚信经营等市场要素,实现多盈利。  相似文献   

20.
[目的]探求有效抑制龙眼组培时外植体褐变的方法。[方法]以龙眼嫩梢茎段为材料,采用多种抗褐变方法,筛选对龙眼组织培养中的外植体褐变抑制效果最好的方法。[结果]采用6.00g/LPVP溶液进行预处理对龙眼外植体褐变的抑制效果最好。当6-BA浓度为1.92~1.93mg/L、KT浓度为0.88~1.93mg/L、NAA浓度为0.05~0.06mg/L时,外植体的褐变程度最轻。添加2.00g/L的PVP和活性炭的培养基对外植体褐变的抑制效果最好。[结论]选取培养基MS+NAA0.05~0.06mg/L+6-BA1.92~1.93mg/L+KT0.88~1.93mg/L+PVP2.00g/L,结合6.00g/LPVP溶液的预处理,对龙眼组织培养时外植体褐变的抑制效果最好。  相似文献   

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