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  • Restriction Enzyme Based Enriched L1Hs Sequencing (REBELseq): A Scalable Technique for Detection of Ta Subfamily L1Hs in the Human Genome.

Restriction Enzyme Based Enriched L1Hs Sequencing (REBELseq): A Scalable Technique for Detection of Ta Subfamily L1Hs in the Human Genome.

G3 (Bethesda, Md.) (2020-03-07)
Benjamin C Reiner, Glenn A Doyle, Andrew E Weller, Rachel N Levinson, Esin Namoglu, Alicia Pigeon, Emilie Dávila Perea, Cynthia Shannon Weickert, Gustavo Turecki, Deborah C Mash, Richard C Crist, Wade H Berrettini
ZUSAMMENFASSUNG

Long interspersed element-1 retrotransposons (LINE-1 or L1) are ∼6 kb mobile DNA elements implicated in the origins of many Mendelian and complex diseases. The actively retrotransposing L1s are mostly limited to the L1 human specific (L1Hs) transcriptional active (Ta) subfamily. In this manuscript, we present REBELseq as a method for the construction of Ta subfamily L1Hs-enriched next-generation sequencing libraries and bioinformatic identification. REBELseq was performed on DNA isolated from NeuN+ neuronal nuclei from postmortem brain samples of 177 individuals and empirically-driven bioinformatic and experimental cutoffs were established. Putative L1Hs insertions passing bioinformatics cutoffs were experimentally validated. REBELseq reliably identified both known and novel Ta subfamily L1Hs insertions distributed throughout the genome. Differences in the proportion of individuals possessing a given reference or non-reference retrotransposon insertion were identified. We conclude that REBELseq is an unbiased, whole genome approach to the amplification and detection of Ta subfamily L1Hs retrotransposons.

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KIMBLE Dounce Gewebehomogenisator, 2 mL complete
Sigma-Aldrich
Anti-NeuN-Antikörper, Klon A60, Alexa Fluor488 konjugiert, clone A60, Chemicon®, from mouse