文档介绍：青岛农业大学
本科生毕业论文
论文题目:烟草T-DNA插入突变体的旁侧序列扩增
专业班级烟草09级01班
姓名(学号)
指导老师姓名
指导老师职称硕士研究生
完成时间 2013年5月28日
摘要: 1
关键词: 1
Abstract: 2
Key words: 2
引言: 3
关键词: 3
4
4
4
4
DNA 纯度、浓度覆分子量的测定 5
5
-PCR 5
6
7
-PCR实验 8
、跑电 9
、纯化DNA 10
. 11
. 12
12
致谢 12
参考文献 12
烟草T-DNA插入突变体的旁侧序列扩增
摘要:通过TALL-PCR技术来分离与已知序列邻近的未知 DNA 序列,利用 3 个根据已知序列设计的嵌套特异性引物分别和简并引物组合进行 PCR 反应,选择恰当退火温度对目标片段进行 PCR 扩增。通过在测序凝胶上电泳条带的比较筛选出不同的表达的基因,并回收这些DNA片段,经扩增后作为探针,在c DNA或基因组DNA库中扫描找到相关基因。应用TALL-PCR技术研究烟草基因功能,能够更好的推动烟草功能基因组学研究。TAIL-PCR 技术作为一种使用技术简单易行,反应高效灵敏,产物特异性高,重复性好,能够在较短的时间内获得目标片段,已经在分子生物学研究领域广泛应用。本文从 TAIL-PCR 技术原理出发,对该技术特异性引物设计、随机引物组合选择、PCR 反应条件等关键性问题进行综述。TAIL-PCR 是一种较为成熟的扩增 T-DNA 插入旁邻序列的技术,它操作相对简单,具有高效性,特异性高,应用非常广泛。
关键词: TALL-PCR技术突变体旁侧序列扩增
Flanking sequence of o T-DNA insertion mutants of amplification
Abstract: through the TALL-PCR technology to separate the unknown DNA sequence adjacent with the known sequence, using 3 nested specific primers according to the known sequence design respectively and degeneracy binations of PCR reaction, selection of appropriate target fragment was amplified by PCR annealing temperature. By sequencing gel electrophoresis bands screened out the different expression genes, and the recovery of these DNA fragments, amplified and used as probe, in C DNA or genomic DNA library scanning to find related genes. Study on the application of TALL-PCR technology in o gene function, can promote the o functional genomics study better. TAIL-PCR technology as a technology is simple, reaction product is high sensitive, high specificity, good reproducibility, can obtain the target fragments in a relatively short period of time, has been widely used in the field of molecular biology research. This article from the TAIL-PCR technology theory, summarizes the technology designed specific primer, binations selected, PCR reaction conditions and other key issu