摘要
<正>以尾叶桉DNA为材料 ,对IdahoRapidcycler扩增仪上用薄壁管进行RAPD扩增的程序进行了优化研究。通过逐步对退火、延伸和变性的温度和时间及总的循环次数设置不同梯度和亚梯度 ,比较不同梯度和亚梯度的扩增结果 ,确定优化程序为 :93℃预变性 1min ;再 40次循环 :93℃变性 2 0s,38℃退火 2 1s,72℃延伸 1min ;最后 72℃延伸 7min。该程序的扩增时间较短 ,具有较好的特异性和重复性。另外 ,薄壁管反应架在反应室的内位比同时扩增的外位效果差 ,扩增时只能利用外位。
Abstract
Inthin walledmicrocentrifugetubes,optimizationofRAPDamplificationprogramforEucalyptusurophyl laS .T .BlakewasmadewithIdahoRapidcycler.Thetemperatureandtimefordenaturation ,annealingandelongation aswellasthenumberofcycleswereindividuallyoptimized .Theprogramachievedwasasfollows :predenaturationat 93℃for1min ;then 40cycles:denaturation 2 0sat93℃ ,annealing2 1sat38℃ ,andelongation 1minat72℃ ; andatlastholding 7minat72℃ .TheprogramcouldresultinsatisfactoryspecificityandrepeatabilityinRAPD amplificationinrelativelyshorttime.Inaddition ,theinnerrowofreactionsinthin walled tubestandperformedweak oremptyinamplificationoflongfragments ,soonlytheouterrowcouldbeusedinpracticaloperation .
甘四明;施季森;白嘉雨.
Idaho Rapidcycler PCR仪上用薄壁管进行RAPD扩增的程序优化[J]. 南京林业大学学报（自然科学版）. 2000, 24(01): 27-31 https://doi.org/10.3969/j.jssn.1000-2006.2000.01.007
GanSiming ShiJisen (LaboratoryofForestGeneticsandGeneEngineeringNanjingForestryUniversity Nanjing 2 10 0 37) BaiJiayu.
Optimization of RAPDAmplification Programin Thin-walled Microcentrifuge TubeswithIdaho Rapidcycler[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2000, 24(01): 27-31 https://doi.org/10.3969/j.jssn.1000-2006.2000.01.007
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