N1-methyladenosine (m1A), N3-methylcytidine (m3C), and N1-methylguanosine (m1G) are common in transfer RNA (tRNA) and tRNA-derived fragments. These modifications alter Watson-Crick base-pairing, and cause pauses or stops during reverse transcription required for most high-throughput RNA sequencing protocols, resulting in inefficient detection of methyl-modified RNAs. Here, we describe a procedure to demethylate RNAs containing m1A, m3C, or m1G using the Escherichia coli dealkylating enzyme AlkB, along with instructions for subsequent processing with widely used protocols for small RNA sequencing.
CITATION STYLE
Hrabeta-Robinson, E., Marcus, E., Cozen, A. E., Phizicky, E. M., & Lowe, T. M. (2017). High-throughput small RNA sequencing enhanced by AlkB-facilitated rna de-methylation (ARM-seq). In Methods in Molecular Biology (Vol. 1562, pp. 231–243). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6807-7_15
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