Vandenbussche Michiel, Zethof Jan, Gerats Tom
UMR 5667 CNRS-INRA-ENS Lyon-Unversité Lyon I, RDP Laboratory, ENS Lyon, Lyon, Cedex, France.
Methods Mol Biol. 2013;1057:251-64. doi: 10.1007/978-1-62703-568-2_18.
The large scale sequencing of insertion element flanking sequences has revolutionized reverse genetics in plant research: Insertion mutants can now simply be identified in silico by BLAST searching the resulting flanking sequence databases. The development of next-generation sequencing technologies has further facilitated the creation of flanking sequence collections derived from entire mutant populations. Here we describe a highly efficient and widely applicable method that we developed to amplify, sequence, and identify dTph1 transposon flanking sequences from a library of 1000 Petunia W138 individuals simultaneously.
现在可以通过对所得侧翼序列数据库进行BLAST搜索，在计算机上简单地鉴定插入突变体。下一代测序技术的发展进一步促进了源自整个突变群体的侧翼序列文库的创建。在这里，我们描述了一种高效且广泛适用的方法，该方法是我们开发的，用于同时从1000个矮牵牛W138个体的文库中扩增、测序和鉴定dTph1转座子侧翼序列。
