10---PCR引物设计(一)汇总课件

1、1第第4章章 生物信息学应用软件生物信息学应用软件4.1 DNAStar4.2 DNAMAN4.3 GeneDoc24.1 DNAStarDNAStar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SeqMan II 序列组装序列组装3安装安装DNAStar应用软件应用软件从从“Bioinformatics Programs”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

2、成动安装，安装完成后从后从“开始开始” “程程序序”中点击中点击“DNAstar”，选择，选择所需程序运行。所需程序运行。44.1.1 EditSeqClick here.开始开始 程序程序 DNAStar EditSeq5Example 4-1试对下面的试对下面的DNA序列进行序列进行EditSeq分析分析.ggcacgaggcgacgaaacaatacttgcctgatttaaacgattataagttttgttaaggaaaaactgtgatcagcttcaaattattttcaaattccgcgtaatctccaagatgtcacacaatggaactttggatgaacattatca

3、gaccgtgagggaattttacgatggtaaatccgtttttattacaggagcgaccggattccttggaaaggcgtacgttgagaaacttgcgtattcgtgtccgggcatagtaagcatttacattttgattcgagacaagaaagggtcgaacactgaagagcgaatgagaaaatatcttgatcagcccattttttcacgaataaaatacgaacatccagaatatttcaaaaaaatcattcccatctctggtgatatcaccgcacccaaactaggattgtgcgatgaagaaagaaatatattaat

4、caacgaggtgtcgatagtgattcactcagccgcttcggtcaaactaaacgatcatttaaaatttactctgaacacgaatgttggaggtacgatgaaagtgttggaactcgttaaagaaatgaaaaacttagcgatgttcgtatatgtgtctacagcttattctaacacaagtcaaagaattttagaagaaaaactttatcctcagtccctaaatctaaacgaaatacagaagttcgctgaagaacattatatattaggtaaagacaacgatgaaatgataaaatttataggtaaccaccc

5、aaacacatatgcatacacaaaagcgctggcagaaaacttagttgctgaagaacacggtgaaataccaacaattatcattaggccttcgattattacagcgagtgctgaagagccagtgcgagggttcgtggatagttggagtggtgcaactgctatggcagcatttgctttgaagggatggaacaacataatgtacagcacaggcgaggagaatattgatctgatacctctagactatgttgtcaaccttacattagtagcgattgctaaatataagcctaccaaagaggtcacagtata

6、tcacgtcacaaccagcgatcttaatcccatttcaatccgaagaatatttataaagctctccgaattcgcatcaaaaaatccaacgagtaatgccgcaccattcgcagcaacgactttattgactaaacagaagccattaataaaactggtcacgtttctcatgcagaccacgccggctttcctagctgacttatggatgaaaacccagaggaaagaagccaaattcgtcaagcaacataatctagtagtgagaagtcgtgatcaactggaattcttcacgtctcagtcgtggttgcttcg

7、ttgcgagcgagcccgtgtgcttagtgcggcactgagcgattcagaccgcgctgtcttccgctgcgatccttccaccatagactgggaccaatatttgccaatatacttcgaaggaatcaacaagcatctgttcaaaaataaattatagtcgattcctaataggcttgtaattgattccaaaaatacttagataacaataactttaatcaactacaatgaaataaatgtaaagtcgtcgtgacctataggataagacgtacggtgcattcgtatatagcgatgcaccggtgttcgaatccc

8、gcaggcggataccaatttttctaatgaaatacgtacttaacaaatgttcacgattgacttccgcggtgaaggaacaacatcgtgtaataaaaatcaaacccgcaaaattataatttgcgtaattactggtggtaggacctcttgtgagtccgcacgggtaggtaccaccactccgtctatttctgccctgaagcagcattgcgtttcggtttgaaaggtggggtagccgtggtaactatactatactatagtaattccaaaattcgctgatttatgttctattagtctgatttagttaaa

9、ccatttctatagttactatagttattacctttcttgtactgcacagagacacggaatgctgttttattttaagttttgagttattatagtttaataaatactgtcatattacataattatttatgtgcaagatgtaatggccatttagatgtgatacgtcctttttacaaaaaaaaaacatgctatatgaaaaaaaaaaaaaaaaaaa6Paste sequencePaste your sequence here.71）Change font sizeClick here.82）Change Layout (1/2)

10、Click here.9Change Layout (2/2)Then click here.Click here.10The changed layout113）Change to UPPERCASEClick here.12Change to uppercaseClick here.13The changed letters144）Find ORF将光标放到第将光标放到第1个碱基上个碱基上Click here.15Find ORFClick here.16The found ORF175）Translate DNAClick here.18Result of “translate DNA”

11、196）Select nucleotides204.1.2 PrimerSelect开始开始 程序程序 DNAStar PrimerSelectClick here.21Example 4-2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-strand Y effector。221）创建）创建PrimerSelect文件文件Click here.23Enter sequenceClick here.24Enter sequence1) Click here.2) Click here.3) Click here

12、.25Block view26Sequence viewClick here.272）定义引物特点和位置）定义引物特点和位置Click here.28Primer characteristics使用默认值，点击使用默认值，点击“OK”。29Primer locations (1/4)Click here.30Primer locations (2/4)Click here.Select “Upper and lower primer ranges”.31Primer locations (3/4)Click here.输入相应数值输入相应数值32Primer locations (4/4)33

13、3）寻找引物对）寻找引物对Click here.34List of primers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Click here.Click here.36Alternate products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）。37Alternate products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40Amplification summary41Composition summary (1/2)Click here.42C