Discover more with SMRT epigenetics
By pairing high throughput with long reads, the PacBio systems offer scalable solutions for assessing CpG methylation in eukaryotic genomes. When using Single Molecule, Real-Time (SMRT) sequencing, genome-wide regional methylation information is generated with no additional sample preparation. For targeted applications, SMRT bisulfite sequencing marries bisulfite samples with highly multiplexed, quantitative long-read sequencing, accurately measuring CpG methylation across ~1.5 kb regions.
From DNA to epigenetic information
- Single library type required (20 kb library prep protocol)
- Size-selection options enrich for longest inserts
- Recommended library inputs of 5 µg of gDNA
- Library automation supported
- For SMRT bisulfite sequencing, generate SMRTbell libraries from long amplicons (1–2 kb) following bisulfite treatment
SMRT sequencing with PacBio systems
- Take advantage of the Sequel system to reduce project costs and generate 7× more reads compared with the PacBio RS II
- Achieve ~10 kb average read lengths, with some reads as long as 60 kb Average read lengths 10–15 kb
Data analysis with SMRT Analysis and PacBio DevNet
- Identify hypo- and hyper-methylated CpG regions in eukaryotic genomes with > 20× coverage
- Detect CpG methylation across 1.5 kb regions with SMRT bisulfite sequencing
PacBio sequencing was used to characterize the cancer genomes of both drug-sensitive and drug-resistant PC-9 cells. Large-scale changes in the methylation status were observed, allowing researchers to explore the function of methylation on drug sensitivity.3
SMRT Sequencing for eukaryotic epigenetics
- Fahrner, J.A. and Bjornson, H.T. (2014) Mendelian disorders of the epigenetic machinery: tipping the balance of chromatin states. Annual Reviews Genomics and Human Genetics. 15, 269-293.
- Baylin, S.B. and Herman J.G. (2000) DNA hypermethylation in tumorigenesis: epigenetics joins genetics. Trends in Genetics. 16(4),168–174.
- Korlach, J., et al., (2015, February) Whole genome sequencing and epigenome characterization of cancer cells using the PacBio platform. Poster presented at Advances in Genome Biology and Technology., Marco Island, FL.