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• W1987291027 endingPage "e10168" @default.
• W1987291027 startingPage "e10168" @default.
• W1987291027 abstract "Transposable elements (such as the P-element and piggyBac) have been used to introduce thousands of transgenic constructs into the Drosophila genome. These transgenic constructs serve many roles, from assaying gene/cell function, to controlling chromosome arm rearrangement. Knowing the precise genomic insertion site for the transposable element is often desired. This enables identification of genomic enhancer regions trapped by an enhancer trap, identification of the gene mutated by a transposon insertion, or simplifying recombination experiments. The most commonly used transgene mapping method is inverse PCR (iPCR). Although usually effective, limitations with iPCR hinder its ability to isolate flanking genomic DNA in complex genomic loci, such as those that contain natural transposons. Here we report the adaptation of the splinkerette PCR (spPCR) method for the isolation of flanking genomic DNA of any P-element or piggyBac. We report a simple and detailed protocol for spPCR. We use spPCR to 1) map a GAL4 enhancer trap located inside a natural transposon, pinpointing a master regulatory region for olfactory neuron expression in the brain; and 2) map all commonly used centromeric FRT insertion sites. The ease, efficiency, and efficacy of spPCR could make it a favored choice for the mapping of transposable element in Drosophila." @default.
• W1987291027 created "2016-06-24" @default.
• W1987291027 creator A5019791073 @default.
• W1987291027 creator A5052084166 @default.
• W1987291027 date "2010-04-13" @default.
• W1987291027 modified "2023-10-16" @default.
• W1987291027 title "Splinkerette PCR for Mapping Transposable Elements in Drosophila" @default.
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• W1987291027 doi "https://doi.org/10.1371/journal.pone.0010168" @default.
• W1987291027 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2854151" @default.
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• W1987291027 hasPublicationYear "2010" @default.
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