Single-locus enrichment without amplification for sequencing and direct detection of epigenetic modifications
| Autor: | Jeremiah Hanes, Regina Lam, Stephen Turner, Paul J. Hagerman, Thang Pham, Jun Yi Wang, Erick Loomis, John Eid, Evan Adams, Jun Yin |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Plant Biology & Botany Biology Cell Line Epigenesis Genetic 03 medical and health sciences chemistry.chemical_compound Fragile X Mental Retardation Protein Tandem repeat Genetics Targeted enrichment Humans Epigenetics Allele FMR1 Molecular Biology 0604 Genetics Epigenetic modification Tandem repeats General Medicine Methylation Sequence Analysis DNA DNA Methylation Human genetics Single molecule sequencing 030104 developmental biology chemistry Tandem Repeat Sequences Fragile X Syndrome DNA methylation Female DNA |
| Zdroj: | Molecular genetics and genomics : MGG. 291(3) |
| ISSN: | 1617-4623 |
| Popis: | A gene-level targeted enrichment method for direct detection of epigenetic modifications is described. The approach is demonstrated on the CGG-repeat region of the FMR1 gene, for which large repeat expansions, hitherto refractory to sequencing, are known to cause fragile X syndrome. In addition to achieving a single-locus enrichment of nearly 700,000-fold, the elimination of all amplification steps removes PCR-induced bias in the repeat count and preserves the native epigenetic modifications of the DNA. In conjunction with the single-molecule real-time sequencing approach, this enrichment method enables direct readout of the methylation status and the CGG repeat number of the FMR1 allele(s) for a clonally derived cell line. The current method avoids potential biases introduced through chemical modification and/or amplification methods for indirect detection of CpG methylation events. |
| Databáze: | OpenAIRE |
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