Reduced representation bisulfite sequencing information
Methylation process
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Reduced representation bisulfite sequencing (RRBS) is an efficient and high-throughput technique for analyzing the genome-wide methylation profiles on a single nucleotide level. It combines restriction enzymes and bisulfite sequencing to enrich for areas of the genome with a high CpG content. Due to the high cost and depth of sequencing to analyze methylation status in the entire genome, Meissner et al. developed this technique in 2005 to reduce the amount of nucleotides required to sequence to 1% of the genome.[1] The fragments that comprise the reduced genome still include the majority of promoters, as well as regions such as repeated sequences that are difficult to profile using conventional bisulfite sequencing approaches.[2]
An outline of the protocol for reduced representation bisulfite sequencing
^Alexander Meissner, Andreas Gnirke, George W. Bell, Bernard Ramsahoye, Eric S. Lander and Rudolf Jaenisch. 2005. "Reduced representation bisulfite sequencing for comparative high-resolution DNA methylation analysis". Nucleic Acids Res. 33(18):5868-77
^Gu H, Smith ZD, Bock C, Boyle P, Gnirke A, Meissner A. 2011. "Preparation of reduced representation bisulfite sequencing libraries for genome-scale DNA methylation profiling." Nat Protoc. 6(4):468-81. doi:10.1038/nprot.2010.190.
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yielded discoveries about the potential of certain chemicals such as bisulfite and aminopurine to change certain bases in a gene. This research continued...
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