Enhanced 5-methylcytosine detection in single-molecule, real-time sequencing via Tet1 oxidation.

ABSTRACT:

Background

DNA methylation serves as an important epigenetic mark in both eukaryotic and prokaryotic organisms. In eukaryotes, the most common epigenetic mark is 5-methylcytosine, whereas prokaryotes can have 6-methyladenine, 4-methylcytosine, or 5-methylcytosine. Single-molecule, real-time sequencing is capable of directly detecting all three types of modified bases. However, the kinetic signature of 5-methylcytosine is subtle, which presents a challenge for detection. We investigated whether conversion of 5-methylcytosine to 5-carboxylcytosine using the enzyme Tet1 would enhance the kinetic signature, thereby improving detection.

Results

We characterized the kinetic signatures of various cytosine modifications, demonstrating that 5-carboxylcytosine has a larger impact on t

SUBMITTER: Clark TA 

PROVIDER: S-EPMC3598637 | biostudies-literature | 2013 Jan

REPOSITORIES: biostudies-literature