Document Detail

Positional cloning by fast-track SNP-mapping in Drosophila melanogaster.
MedLine Citation:
PMID:  18948975     Owner:  NLM     Status:  MEDLINE    
Positional cloning of chemically induced mutations is the rate-limiting step in forward genetic screens in Drosophila. Single-nucleotide polymorphisms (SNPs) are useful markers to locate a mutated region in the genome. Here, we provide a protocol for high-throughput, high-resolution SNP mapping that enables rapid and cost-effective positional cloning in Drosophila. In stage 1 of the protocol, we use highly multiplexed tag-array mini-sequencing assays to map mutations to an interval of 1-2 Mb. In these assays, SNPs are genotyped by primer extension using fluorescently labeled dideoxy-nucleotides. Fluorescent primers are captured and detected on a microarray. In stage 2, we selectively isolate recombinants within the identified 1-2 Mb interval for fine mapping of mutations to about 50 kb. We have previously demonstrated the applicability of this protocol by mapping 14 muscle morphogenesis mutants within 4 months, which represents a significant acceleration compared with other commonly used mapping strategies that may take years.
Frank Schnorrer; Annika Ahlford; Doris Chen; Lili Milani; Ann-Christine Syvänen
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Nature protocols     Volume:  3     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2008  
Date Detail:
Created Date:  2008-10-24     Completed Date:  2009-01-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1751-65     Citation Subset:  IM    
Research Institute of Molecular Pathology, Dr Bohr-Gasse 7, A-1030 Vienna, Austria.
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MeSH Terms
Chromosome Mapping / methods*
Cloning, Molecular / methods*
Drosophila melanogaster / embryology,  genetics*
Embryo, Nonmammalian
Genetic Markers
Nucleic Acid Hybridization
Oligonucleotide Array Sequence Analysis
Polymerase Chain Reaction
Polymorphism, Single Nucleotide*
Recombination, Genetic
Reg. No./Substance:
0/Genetic Markers

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