| Chapter title |
Genome-Wide Profiling of DNA Methyltransferases in Mammalian Cells
|
|---|---|
| Chapter number | 9 |
| Book title |
CpG Islands
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7768-0_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7767-3, 978-1-4939-7768-0
|
| Authors |
Massimiliano Manzo, Christina Ambrosi, Tuncay Baubec |
| Abstract |
Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) is currently the method of choice to determine binding sites of chromatin-associated factors in a genome-wide manner. Here, we describe a method to investigate the binding preferences of mammalian DNA methyltransferases (DNMT) based on ChIP-seq using biotin-tagging. Stringent ChIP of DNMT proteins based on the strong interaction between biotin and avidin circumvents limitations arising from low antibody specificity and ensures reproducible enrichment. DNMT-bound DNA fragments are ligated to sequencing adaptors, amplified and sequenced on a high-throughput sequencing instrument. Bioinformatic analysis gives valuable information about the binding preferences of DNMTs genome-wide and around promoter regions. This method is unconventional due to the use of genetically engineered cells; however, it allows specific and reliable determination of DNMT binding. |
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| Canada | 1 | 8% |
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Demographic breakdown
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Mendeley readers
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|---|---|---|
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Demographic breakdown
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|---|---|---|
| Researcher | 4 | 31% |
| Student > Bachelor | 2 | 15% |
| Other | 1 | 8% |
| Student > Ph. D. Student | 1 | 8% |
| Student > Doctoral Student | 1 | 8% |
| Other | 2 | 15% |
| Unknown | 2 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 8 | 62% |
| Neuroscience | 1 | 8% |
| Chemistry | 1 | 8% |
| Medicine and Dentistry | 1 | 8% |
| Unknown | 2 | 15% |