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| Status |
Public on Oct 09, 2018 |
| Title |
Bisulfite-free, nano-scale analysis of 5-hydroxymethylcytosine at single base resolution |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Methylation profiling by high throughput sequencing
Other
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| Summary |
High-resolution detection of genome-wide 5-hydroxymethylcytosine (5hmC) sites of small-scale samples represents a continuous challenge. Here, we present CATCH-seq, a bisulfite-free, base-resolution method for the genome-wide detection of 5hmC. CATCH-seq is based on selective 5hmC oxidation, labeling and subsequent C-to-T transition during PCR. Applications of CATCH-seq to nano-scale DNA samples reveal previously underappreciated non-CG 5hmCs in the hESC genome and base-resolution hydroxymethylome in human cell-free DNA.
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| Overall design |
We developed a method named "CATCH-Seq" , a bisulfite-free, base-resolution method for the genome-wide detection of 5hmC. Our method is based on: (1) selective oxidation of 5hmC to 5fC ; (2) subsequent labeling of the newly generated 5fC; and (3) 5fC labeling adduct caused C-to-T transition during DNA amplification.We then applied CATCH-seq to 100 ng, 50 ng and 10 ng genomic DNA isolated from hESCs, respectively. To further demonstrate the utility of CATCH-seq in analyzing precious clinical samples, we next applied this approach to sequence the hydroxymethylome of cell-free DNA (cfDNA).
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| Contributor(s) |
Zeng H, He B, Xia B, Yi C |
| Citation(s) |
30278133 |
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| Submission date |
Mar 19, 2018 |
| Last update date |
Mar 27, 2019 |
| Contact name |
Bo He |
| Organization name |
Peking University
|
| Department |
School of Life Science
|
| Street address |
Yiheyuan Road 5th.
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| City |
Beijing |
| State/province |
Beijing |
| ZIP/Postal code |
100871 |
| Country |
China |
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| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
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| Samples (18)
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| Relations |
| BioProject |
PRJNA439006 |
| SRA |
SRP136030 |
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