RevGenUK, the next generation: establishing a TILLING boutique for a UK-based reverse genetics community resource
Lead Research Organisation:
Earlham Institute
Department Name: UNLISTED
Abstract
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Technical Summary
TILLING (Targeting Induced Local Lesions in Genomes) is a non-GM reverse genetics technology for discovering, characterising and exploiting genetic variation, based on mutation detection. Reverse genetics is a strategy whereby plants carrying mutations in target genes are identified and then phenotyped, in order to determine the function of those genes. RevGenUK is a global TILLING service previously established for Medicago truncatula and Brassica rapa. The aim of this project is to expand the species range and to implement TILLING-by-sequencing as the strategy for mutation detection.
Recent advances in next generation sequencing (NGS) throughput, the ability to barcode and pool samples prior to sequencing, and increased read-lengths have made TILLING-by-sequencing a practical alternative to conventional TILLING. In addition the use of exome capture approaches allow the targeted sequencing of regions of interest within the genome. Reducing the fraction of the genome to be sequenced, enables increased multiplexing of samples, and a reduction in both cost and analysis time. Over one thousand mutagenised lines of B. rapa will undergo exome capture, resequencing and analysis to detect mutations within the captured gene regions. These mutations will be displayed within a web-accessible database enabling community access to these materials. These developments will enable RevGenUK to provide a high throughput, efficient and cost effective service at the cutting edge of reverse genetics technologies.
Recent advances in next generation sequencing (NGS) throughput, the ability to barcode and pool samples prior to sequencing, and increased read-lengths have made TILLING-by-sequencing a practical alternative to conventional TILLING. In addition the use of exome capture approaches allow the targeted sequencing of regions of interest within the genome. Reducing the fraction of the genome to be sequenced, enables increased multiplexing of samples, and a reduction in both cost and analysis time. Over one thousand mutagenised lines of B. rapa will undergo exome capture, resequencing and analysis to detect mutations within the captured gene regions. These mutations will be displayed within a web-accessible database enabling community access to these materials. These developments will enable RevGenUK to provide a high throughput, efficient and cost effective service at the cutting edge of reverse genetics technologies.
Planned Impact
unavailable
