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PMID: 17645438 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Indel arrays: an affordable alternative for genotyping.

The Plant journal : for cell and molecular biology ·Vol. 51 ·No. 4 ·2007-08-00 ·Pages 727-37

Salathia N, Lee HN, Sangster TA, Morneau K, Landry CR, Schellenberg K, Behere AS, Gunderson KL, Cavalieri D, Jander G, Queitsch C

Abstract

Natural variation and induced mutations are important resources for gene discovery and the elucidation of genetic circuits. Mapping such polymorphisms requires rapid and cost-efficient methods for genome-wide genotyping. Here we report the development of a microarray-based method that assesses 240 unique markers in a single hybridization experiment at a cost of less than US$50 in materials per line. Our genotyping array is built with 70-mer oligonucleotide elements representing insertion/deletion (indel) polymorphisms between the Arabidopsis thaliana accessions Columbia-0 (Col) and Landsberg erecta (Ler). These indel polymorphisms are recognized with great precision by comparative genomic hybridization, eliminating the need for array replicates and complex statistical analysis. Markers are present genome-wide, with an average spacing of approximately 500 kb. PCR primer information is provided for all array indels, allowing rapid single-locus inquiries. Multi-well chips allow groups of 16 lines to be genotyped in a single experiment. We demonstrate the utility of the array for accurately mapping recessive mutations, RIL populations and mixed genetic backgrounds from accessions other than Col and Ler. Given the ease of use of shotgun sequencing to generate partial genomic sequences of unsequenced species, this approach is readily transferable to non-model organisms.

MeSH Terms
Arabidopsis/genetics Gene Deletion Genetic Markers/genetics Genome, Plant Genotype Mutagenesis, Insertional Oligonucleotide Array Sequence Analysis/methods Polymorphism, Genetic/genetics Species Specificity
Chemicals
Genetic Markers
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Salathia Neeraj
FAS Center for Systems Biology, Harvard University, Cambridge, MA 02138, USA.
Lee Hana N
Sangster Todd A
Morneau Keith
Landry Christian R
Schellenberg Kurt
Behere Aditi S
Gunderson Kevin L
Cavalieri Duccio
Jander Georg
Queitsch Christine
Article Info
Journal
The Plant journal : for cell and molecular biology
Abbr.
Plant J
ISSN
0960-7412
Published
2007-08-00
Epub
2007-00-20
Pages
727-37
Language
English
Region
England
NLM ID
9207397
Subset
IM
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