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Fig. 1 | Mobile DNA

Fig. 1

From: SeqURE – a new copy-capture based method for sequencing of unknown Retroposition events

Fig. 1

Principle of the method. Genomic DNA is digested by a mixture of restrictases that do not cut inside Alu element. After adapter ligation and linear amplification with AluYa5 or AluYb8 specific primer the copy of Alu 5′ flank is captured by streptavidin coated magnetic beads. This procedure is repeated for the 3′ flank of Alu. Captured linear amplification products are used in two stage exponential PCR (1st PCR + indexing PCR) to generate libraries for sequencing on Illumina. Remaining template DNA is used for validation in locus-specific PCR. UMI – Unique Molecular Identifier, dAlu – part of Alu repeat, i7 and i5 standard Illumina Nextera sample barcodes. Blue circles indicate biotin attached to the Alu specific primers and dCTP

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