TY  - JOUR
TI  - Identification of Methylated Deoxyadenosines in Genomic DNA by dA6m DNA Immunoprecipitation
AU  - Koziol, Magdalena  J.
AU  - Bradshaw, Charles  R.
AU  - Allen, George  E.
AU  - Costa, Ana S. H.
AU  - Frezza, Christian  
VL  - 6
IS  - 21
PY  - 2016
DA  - 2016/11/05
SP  - e1990
C1  - Bio-protocol 2016;6:e1990
DO  - 10.21769/BioProtoc.1990
UR  - https://doi.org/10.21769/BioProtoc.1990
AB  - dA6m DNA immunoprecipitation followed by deep sequencing (DIP-Seq) is a key tool in identifying and studying the genome-wide distribution of N6-methyldeoxyadenosine (dA6m). The precise function of this novel DNA modification remains to be fully elucidated, but it is known to be absent from transcriptional start sites and excluded from exons, suggesting a role in transcriptional regulation (Koziol et al., 2015). Importantly, its existence suggests that DNA might be more diverse than previously believed, as further DNA modifications might exist in eukaryotic DNA (Koziol et al., 2015). This protocol describes the method to perform dA6m DNA immunoprecipitation (DIP), as was applied to characterize the first dA6m methylome analysis in higher eukaryotes (Koziol et al., 2015). In this protocol, we describe how genomic DNA is isolated, fragmented and then DNA containing dA6m is pulled down with an antibody that recognizes dA6m in genomic DNA. After subsequent washes, DNA fragments that do not contain dA6m are eliminated, and the dA6m containing fragments are eluted from the antibody in order to be processed further for subsequent analyses.
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False