Virtually all methods for reading the sequence of bases in DNA rely on the ability to denature double-stranded DNA into single strands and then use Watson-Crick base-pairing rules to hybridize the strands with high specificity to another DNA primer or probe. However, nature frequently uses an alternative method, reading the sequence information directly from double-stranded DNA using sequence-specific DNA-binding proteins. Here we describe methods for the construction and testing of sequence probes based on engineered zinc finger DNA-binding proteins. Background is reduced using split-reporter molecules, and signal is amplified using enzymatic reporters. The resulting sequence-enabled reassembly (SEER) probes can be configured to detect DNA sequence (genetic) or DNA methylation (epigenetic) information. © 2010 Springer Science+Business Media, LLC.
CITATION STYLE
Porter, J. R., Lockwood, S. H., Segal, D. J., & Ghosh, I. (2010). Seeing genetic and epigenetic information without DNA denaturation using sequence-enabled reassembly (SEER). Methods in Molecular Biology, 649, 365–382. https://doi.org/10.1007/978-1-60761-753-2_23
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