Table_3_TDNAscan: A Software to Identify Complete and Truncated T-DNA Insertions.xlsx (10.13 kB)
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Table_3_TDNAscan: A Software to Identify Complete and Truncated T-DNA Insertions.xlsx

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posted on 05.08.2019, 13:15 authored by Liang Sun, Yinbing Ge, J. Alan Sparks, Zachary T. Robinson, Xiaofei Cheng, Jiangqi Wen, Elison B. Blancaflor

Transfer (T)-DNA insertions in mutants isolated from forward genetic screens are typically identified through thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR), inverse PCR, or plasmid rescue. Despite the popularity and success of these methods, they have limited capabilities, particularly in situations in which the T-DNA is truncated. Here, we present a next generation sequencing (NGS)-based platform to facilitate the identification of complete and truncated T-DNA insertions. Our method enables the detection of the corresponding T-DNA insertion orientation and zygosity as well as insertion annotation. This method, called TDNAscan, was developed to be an open source software. We expect that TDNAscan will be a valuable addition to forward genetics toolkits because it provides a solution to the problem of causal gene identification, particularly genes disrupted by truncated T-DNA insertions. We present a case study in which TDNAscan was used to determine that the recessive Arabidopsis thaliana hypersensitive to latrunculin B (hlb3) mutant isolated in a forward genetic screen of T-DNA mutagenized plants encodes a class II FORMIN.

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