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
Whole genome sequencing and epigenome characterization of cancer cells
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.