Table S1. Primers for quality control qPCR from Human transposon insertion profiling by sequencing (TIPseq) to map LINE-1 insertions in single cells
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https://rs.figshare.com/articles/Table_S1_Primers_for_quality_control_qPCR_from_Human_transposon_insertion_profiling_by_sequencing_TIPseq_to_map_LINE-1_insertions_in_single_cells/11449998
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Long interspersed element-1 (LINE-1, L1) sequences, which comprise about 17% of human genome, are the product of one of the most active types of mobile DNAs in modern humans. LINE-1 insertion alleles can cause inherited and de novo genetic diseases, and LINE-1-encoded proteins are highly expressed in some cancers. Genome-wide LINE-1 mapping in single cells could be useful for defining somatic and germline retrotransposition rates, and for enabling studies to characterize tumour heterogeneity, relate insertions to transcriptional and epigenetic effects at the cellular level or describe cellular phylogenies in development. Our laboratories have reported a genome-wide LINE-1 insertion site mapping method for bulk DNA, named transposon insertion profiling by sequencing (TIPseq). There have been significant barriers applying LINE-1 mapping to single cells, due to the chimeric artefacts and features of repetitive sequences. Here, we optimize a modified TIPseq protocol and show its utility for LINE-1 mapping in single lymphoblastoid cells. Results from single-cell TIPseq experiments compare well to known LINE-1 insertions found by whole-genome sequencing and TIPseq on bulk DNA. Among the several approaches we tested, whole-genome amplification by multiple displacement amplification followed by restriction enzyme digestion, vectorette ligation and LINE-1-targeted PCR had the best assay performance.This article is part of the discussion meeting issue ‘Crossroads between transposons and gene regulation’.
提供机构:
The Royal Society
创建时间:
2019-12-24



