Jan 22, 2016

Tagmentation-Based Mapping (TagMap) of Mobile DNA Genomic Insertion Sites

BioRxiv : the Preprint Server for Biology
David L. Stern

Abstract

Multiple methods have been introduced over the past 30 years to identify the genomic insertion sites of transposable elements and other DNA elements that integrate into genomes. However, each of these methods suffer from limitations that can frustrate attempts to map multiple insertions in a single genome and to map insertions in genomes of high complexity that contain extensive repetitive DNA. I introduce a new method for transposon mapping that is simple to perform, can accurately map multiple insertions per genome, and generates long sequence reads that facilitate mapping to complex genomes. The method, called TagMap, for Tagmentation-based Mapping, relies on a modified Tn5 tagmentation protocol with a single tagmentation adaptor followed by PCR using primers specific to the tranposable element and the adaptor sequence. Several minor modifications to normal tagmentation reagents and protocols allow easy and rapid preparation of TagMap libraries. Short read sequencing starting from the adaptor sequence generates oriented reads that flank and are oriented toward the transposable element insertion site. The convergent orientation of adjacent reads at the insertion site allows straightforward prediction of the precise insertion ...Continue Reading

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Mentioned in this Paper

Genome
Genomic Stability
DNA Sequence - Cloning Site
Site
Genomics
Sequencing
Oligonucleotide Primers
Genes, Jumping
transposon/insertion element
Tn5 transposase

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