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).
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