Selection of efficient Taq DNA polymerase to optimize T-DNA genotyping method for rapid detection of mutant Arabidopsis thaliana plants.
Selection of efficient Taq DNA polymerase to optimize T-DNA genotyping method for rapid detection of mutant Arabidopsis thaliana plants.
复制标题
选择高效Taq DNA聚合酶来优化T-DNA基因分型方法，用于快速检测突变拟南芥植物。
DOI：
10.1007/s12668-016-0253-6
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发表时间：
2016
期刊：
影响因子：
3
通讯作者：
Shakirov,EugeneV
中科院分区：
文献类型：
--
作者：
Chastukhina,InnaB;Nigmatullina,LiliiaR;Valeeva,LiaR;Shakirov,EugeneV
植物含有参与磷代谢、端粒生物学和其他细胞过程的各种动物基因的同源物。与许多其他多细胞生物的实验相比，模式植物拟南芥的研究利用了较短的世代时间和不断增加的遗传和转基因工具库，包括大量的 T-DNA 敲除和激活系。数以千计的公开转基因拟南芥属植物的可用性为解决许多重要的生物学问题提供了独特的机会。然而，从生物库存分配中心提供的种子池中鉴定单个 T-DNA 突变植物仍然是一个费力且耗时的过程。在这里，我们比较了一些常用于常规 PCR 基因分型的商业 Taq DNA 聚合酶，以确定最适合 T-DNA 突变植物基因分型的单一聚合酶。我们的数据表明，Emerald Amp GT PCR Master Mix 提供了最可靠、快速和简单的 DNA 基因分型工具，用于确定 T-DNA 插入的存在并确定个体 A 是否存在。拟南芥的突变等位基因是杂合的或纯合的。
Plants harbor homologues of various animal genes involved in phosphorus metabolism, telomere biology and other cellular processes. Compared to experiments with many other multicellular organisms, research in the model plantArabidopsis thalianatakes advantage of short generation time and an ever increasing arsenal of genetic and transgenic tools, including large collections of T-DNA knockout and activation lines. The availability of thousands of publicly available transgenicArabidopsislines provides a unique opportunity to address a number of important biological questions. However, identification of individual T-DNA mutant plants from a pool of seeds provided by a biological stock distribution center remains a laborious and time-consuming procedure. Here we compared a number of commercial Taq DNA polymerases commonly used for routine PCR genotyping to identify a single polymerase most suitable for genotyping T-DNA mutant plants. Our data indicate that Emerald Amp GT PCR Master Mix provides the most reliable, quick and simple DNA genotyping tool to determine the presence of a T-DNA insertion and to establish whether an individualA. thalianaplant is heterozygous or homozygous for the mutant allele.