DNA Adenine Methyltransferase Identification (DamID) Coupled to Next Generation Sequencing For Profiling Lamina Associated Domains

Purpose: To compare the interaction of various proteins (in particular Lap2b and Lamin A) at the nuclear periphery with the Lamina Associated Domains (LADs) Results: Using an optimized data analysis workflow (LADetector, available at https://github.com/thereddylab/LADetector), We found near identical sequences within the genome-wide maps indicating that LAP2β and Lamin A, and hence LaminB1, are in close proximity to all LADs, suggesting that these proteins are not involved in discriminating subsets of LADs. LAD profiles, of MEFs, corresponding to various proteins at the nuclear periphery (LMNA, Lap2b) were generated following the DamID protocol (L.Guelen et al, Nature 2008), library randomization and sequencing preparation using Illumina Paired End Sequencing Kit and sequenced on the Illumina HiSeq 2000.