PCR引物设计(一)汇总课件.ppt

1第第4章章生物信息学应用软件生物信息学应用软件4.1DNAStar4.2DNAMAN4.3GeneDoc24.1DNAStarDNAStar的主要功能：的主要功能：EditSeq序列编辑序列编辑GeneQuest基因搜索基因搜索MapDraw图谱绘制图谱绘制MegAlign序列比对序列比对PrimerSelect引物设计引物设计Protean蛋白分析蛋白分析SeqManII序列组装序列组装3安装安装DNAStar应用软件应用软件从从“BioinformaticsPrograms”文件文件夹中找夹中找Dnastar可可执行文件，双击启执行文件，双击启动安装，安装完成动安装，安装完成后从后从“开始开始”“程序程序”中点击中点击“DNAstar”，选择选择所需程序运行。所需程序运行。44.1.1EditSeqClickhere.开始开始程序程序 DNAStarEditSeq5Example4-1试对下面的试对下面的DNA序列进行序列进行EditSeq分析分析.ggcacgaggcgacgaaacaatacttgcctgatttaaacgattataagttttgttaaggaaaaactgtgatcagcttcaaattattttcaaattccgcgtaatctccaagatgtcacacaatggaactttggatgaacattatcagaccgtgagggaattttacgatggtaaatccgtttttattacaggagcgaccggattccttggaaaggcgtacgttgagaaacttgcgtattcgtgtccgggcatagtaagcatttacattttgattcgagacaagaaagggtcgaacactgaagagcgaatgagaaaatatcttgatcagcccattttttcacgaataaaatacgaacatccagaatatttcaaaaaaatcattcccatctctggtgatatcaccgcacccaaactaggattgtgcgatgaagaaagaaatatattaatcaacgaggtgtcgatagtgattcactcagccgcttcggtcaaactaaacgatcatttaaaatttactctgaacacgaatgttggaggtacgatgaaagtgttggaactcgttaaagaaatgaaaaacttagcgatgttcgtatatgtgtctacagcttattctaacacaagtcaaagaattttagaagaaaaactttatcctcagtccctaaatctaaacgaaatacagaagttcgctgaagaacattatatattaggtaaagacaacgatgaaatgataaaatttataggtaaccacccaaacacatatgcatacacaaaagcgctggcagaaaacttagttgctgaagaacacggtgaaataccaacaattatcattaggccttcgattattacagcgagtgctgaagagccagtgcgagggttcgtggatagttggagtggtgcaactgctatggcagcatttgctttgaagggatggaacaacataatgtacagcacaggcgaggagaatattgatctgatacctctagactatgttgtcaaccttacattagtagcgattgctaaatataagcctaccaaagaggtcacagtatatcacgtcacaaccagcgatcttaatcccatttcaatccgaagaatatttataaagctctccgaattcgcatcaaaaaatccaacgagtaatgccgcaccattcgcagcaacgactttattgactaaacagaagccattaataaaactggtcacgtttctcatgcagaccacgccggctttcctagctgacttatggatgaaaacccagaggaaagaagccaaattcgtcaagcaacataatctagtagtgagaagtcgtgatcaactggaattcttcacgtctcagtcgtggttgcttcgttgcgagcgagcccgtgtgcttagtgcggcactgagcgattcagaccgcgctgtcttccgctgcgatccttccaccatagactgggaccaatatttgccaatatacttcgaaggaatcaacaagcatctgttcaaaaataaattatagtcgattcctaataggcttgtaattgattccaaaaatacttagataacaataactttaatcaactacaatgaaataaatgtaaagtcgtcgtgacctataggataagacgtacggtgcattcgtatatagcgatgcaccggtgttcgaatcccgcaggcggataccaatttttctaatgaaatacgtacttaacaaatgttcacgattgacttccgcggtgaaggaacaacatcgtgtaataaaaatcaaacccgcaaaattataatttgcgtaattactggtggtaggacctcttgtgagtccgcacgggtaggtaccaccactccgtctatttctgccctgaagcagcattgcgtttcggtttgaaaggtggggtagccgtggtaactatactatactatagtaattccaaaattcgctgatttatgttctattagtctgatttagttaaaccatttctatagttactatagttattacctttcttgtactgcacagagacacggaatgctgttttattttaagttttgagttattatagtttaataaatactgtcatattacataattatttatgtgcaagatgtaatggccatttagatgtgatacgtcctttttacaaaaaaaaaacatgctatatgaaaaaaaaaaaaaaaaaaa6PastesequencePasteyoursequencehere.71）ChangefontsizeClickhere.82）ChangeLayout(1/2)Clickhere.9ChangeLayout(2/2)Thenclickhere.Clickhere.10Thechangedlayout113）ChangetoUPPERCASEClickhere.12ChangetouppercaseClickhere.13Thechangedletters144）FindORF将将光光标标放放到到第第1个个碱碱基基上上Clickhere.15FindORFClickhere.16ThefoundORF175）TranslateDNAClickhere.18Resultof“translateDNA”196）Selectnucleotides204.1.2PrimerSelect开始开始程序程序DNAStarPrimerSelectClickhere.21Example4-2以克隆载体以克隆载体pBR322为模板为模板DNA序列，设计引序列，设计引物用来扩增位于物用来扩增位于2013-2169位的位的H-strandYeffector。221）创建）创建PrimerSelect文件文件Clickhere.23EntersequenceClickhere.24Entersequence1)Clickhere.2)Clickhere.3)Clickhere.25Blockview26SequenceviewClickhere.272）定义引物特点和位置）定义引物特点和位置Clickhere.28Primercharacteristics使用默认值，点击使用默认值，点击“OK”。29Primerlocations(1/4)Clickhere.30Primerlocations(2/4)Clickhere.Select“Upperandlowerprimerranges”.31Primerlocations(3/4)Clickhere.输入相应数值输入相应数值32Primerlocations(4/4)333）寻找引物对）寻找引物对Clickhere.34Listofprimers按住圆圈往按住圆圈往左左移移35对引物的评价对引物的评价Clickhere.Clickhere.36Alternateproducts线条越粗，能扩增线条越粗，能扩增出的产物量越大出的产物量越大（第（第1对引物）对引物）。37Alternateproducts线条不是很粗，扩线条不是很粗，扩增出的产物量较少增出的产物量较少（第（第2对引物）对引物）。38错配点错配点394）查看引物的其他信息）查看引物的其他信息再点击这里再点击这里先选定第一对引物先选定第一对引物40Amplificationsummary41Compositionsummary(1/2)Clickhere.42Compositionsummary(2/2)43Primerselfdimers(1/2)Clickhere.44Primerselfdimers(2/2)45Primerpairdimers(1/2)Clickhere.46Primerpairdimers(2/2)47Primerhairpins(1/2)Clickhere.48Primerhairpins(2/2)