Abstract
Characterization of viral DNA methylation patterns is essential to understand its function in viral pathogenesis. Bisulfite modification, followed by polymerase chain reaction (PCR) and sequencing is the most effective method for the high resolution methylation mapping of viral genomes. Since the bisulfite modification and PCR steps are the most critical ones, an optimized protocol for these two steps is presented, with special attention to potential pitfalls.
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Acknowledgment
D. S. is a recipient of a Cancer Research Fellowship of the Cancer Research Institute (New York)/Concern Foundation (Los Angeles). Raw sequence data of the EBV Cp in Fig. 1 obtained from bisulfite modified C2G6 DNA was kindly provided by Drs. Ferenc Banati and Kalman Szenthe (RT-Europe Nonprofit Research Ltd, Mosonmagyarovar, Hungary).
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Salamon, D. (2017). Analysis of Viral Epigenotypes Using Bisulfite Sequencing: A Detailed Protocol for the Crucial Bisulfite Modification and PCR Amplification Steps. In: Minarovits, J., Niller, H. (eds) Epstein Barr Virus. Methods in Molecular Biology, vol 1532. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6655-4_15
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DOI: https://doi.org/10.1007/978-1-4939-6655-4_15
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Publisher Name: Humana Press, New York, NY
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