The LINE-1 Mobile Element NGS Assay uses primers specifically designed to amplify the 3’ L1H mobile genetic elements in randomly-sheared DNA (Step 1) to detect novel L1H genomic insertions. An L1H 3’-UTR-specific primer (GSP1) is used for a single round of extension into the chromosome (Step 2), followed by ligation of a T-Adapter to generate the template for PCR amplification (Step 3). Two rounds of amplification follow synthesis of the 3’ L1H templates (Steps 4 & 5). The first round uses GSP1 and the Adapter Primer. The second round of PCR uses a nested GSP2 Anchor Primer with the Adapter Primer. A third round of amplification with primers containing the Illumina P5 and P7 adapter sequences (Step 6) prepares the samples for sequencing on the Illumina HiSeq or NextSeq instrument (Step 7).

LINE-1 Mobilization Assay Protocol Overview
Last modified: Sep 11, 2017

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