文章基本信息

标题：High-Efficiency Thermal Asymmetric InterLaced (HE-TAIL) PCR for Amplification of Ds Transposon Insertion Sites in Barley
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作者：Qi Tan, Han ; Singh, Jaswinder
期刊名称：Journal of Plant Molecular Biology and Biotechnology
印刷版ISSN：2146-0302
出版年度：2011
卷号：2
期号：1
页码：9-14
语种：English
出版社：Journal of Plant Molecular Biology and Biotechnology
摘要： Thermal Asymmetric Interlaced PCR (TAIL PCR) has been used in many different species to isolate flanking sequences adjacent to known sequences. This method has always been a challenge in large genome species, therefore alternative methods have been employed to obtain the flanking sequences. However, these methods are expensive and laborious. Here, we have devised a new and improved method to obtain flanking sequences from barley genome. The proposed method is named as the High-efficiency Thermal Asymmetric Interlaced PCR (HE-TAIL PCR). We have introduced a new 15-mer sequence from the green fluorescent protein (GFP). The new primers efficiently generated transposon flanking sequences in the newly generated barleyDs insertion lines as compared to previously reported hiTAIL PCR by Liu and Chen (2007). Using TAIL PCR, minimal manipulation of genomic DNA is required and large number of samples can be performed at the same time, increasing the efficiency of PCR based amplification of flanking sequences. This HE-TAIL PCR method has effectively introduced the many benefits of TAIL PCR into large genome cereals research.
关键词：Activator/ Dissociation (Ac/Ds); Cereals; Flanking sequences; Functional genomics; Gene tagging