Edinburgh, Scotland, United Kingdom - TAIR

Next-generation genetics: mapping and
mutant identification in one step by deep
sequencing
Identification of causative point mutations after forward genetic mutant screens
typically begins with genetic mapping, followed by transformation rescue or
candidate gene sequencing. We present a one step alternative: performing
hundreds of thousands of genotyping assays while sequencing all candidate
genes. This is accomplished by deep sequencing of a pool of F2 progeny
obtained by crossing to a polymorphic strain and does not require prior
knowledge of mapping markers. The application of high-throughput sequencing
shortens the overall time required for genetic mapping from months to weeks
and, importantly, greatly reduces investigator hands-on time. The steps requiring
investigator input are: DNA isolation (1 day), library preparation and validation (4
days), Illumina cluster generation and sequencing (2 days), and data analysis (1
day). Once the mapping population has been established, the present method
therefore allows a single investigator to identify a causative mutation within only
eight working days – approximately an order of magnitude faster than with
conventional methods.
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Wednesday 14:00 - 14:15
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Korbinian Schneeberger1
Stephan Ossowski1
Christa Lanz1
Trine Juul2
Annabeth Høgh
Petersen3
Kåre Lehmann Nielsen3
Jan-Elo Jørgensen2
Detlef Weigel1
Stig Uggerhøj Andersen2
1Department of Molecular
Biology
Max Planck Institute for
Developmental Biology
Spemannstrasse 37-39
D-72076 Tübingen
Germany
2Department of Molecular
Biology
University of Aarhus
Gustav Wieds Vej 10
DK-8000 Aarhus
Denmark
3Department of Life
Sciences
University of Aalborg
Sohngørdsholmsvej 49
DK-9000 Aalborg
Denmark