First International

Epigenomics & Sequencing 2007

Meeting on ‘Chromatin Methylation to Disease Biology & Theranostics’

July 9-10, 2007
The Conference Center at Harvard Medical School
77 Avenue Louis Pasteur, Boston, MA 02115, USA

“A Unique Theme to Combine Chromatin biology and Diseases with Sequencing Chemistry”

Scientific Sessions Starts at 9:00 A.M and Ends at 5:30 P.M on both days.

Scientific Organizing Committee:

	Krishnarao Appasani, PhD., MBA (Chair) Founder & CEO GeneExpression Systems, Inc. Waltham, MA USA
	Bradley E. Bernstein, M.D. Ph.D. Assistant Professor of Pathology Massachusetts General Hospital, Harvard Medical School, Boston, MA USA Title: Chromatin state maps for pluripotent and committed cells
	Laurie Jackson-Grusby, Ph.D. Assistant Professor Pathology Children's Hospital Boston, Harvard Medical School, Boston, MA, USA
	Mukesh Verma, PhD. Program Director of Epidemiology and Genetics National Cancer Institute, National Institutes of Health, Bethesda, MD
	Alex Meissner, Ph.D. Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology Cambridge, MA, USA Title: Sequencing of Epigenome

Key Topics:
Mechanisms of Chromatin in gene regulation
Nuclear dynamics, ChIP on ChIP and Methylation Assays
Parental imprinting and Histone Deacetylation inhibitors as drugs
Epigenetic re-programming in stem cells
Epigenome Sequencing
Epigenetic regulatory processes in diseases & environment
PharmacoEpigenomics

AGENDA/Speakers

Click Here For Agenda

	Keynote Speaker on July 9 Manel Esteller, M.D., Ph.D. Director, Cancer Epigenetics Laboratory Molecular Pathology Program Spanish National Cancer Centre, Madrid, Spain Title: The Epigenomes of Cancer Cells
	Industry Keynote Speaker on July 10 Maithreyan Srinivasan PhD. Director of Amplification and Sequencing Technology 454 Life Sciences 20 Commercial Street Branford, CT  06405 Title: Comprehensive gene regulation studies using 454 Sequencing™

Other Speakers:

Craig Peterson, Ph.D.
Professor, Program in Molecular Medicine
University of Massachusetts Medical School, Worcester, MA, USA

David E. Fisher MD, PhD
Director, Melanoma Program in Medical Oncology &
Professor of Pediatric Hematology/Oncology
Dana-Farber Cancer Institute & Children's Hospital Boston, Harvard Medical School
Boston, MA, USA
Title: Nucleosome Positioning and relevance to disease

Tomas J. Ekström, Ph.D.
Professor
Laboratory for Molecular Development and Tumor Biology
Karolinska Institutet & Karolinska Hospital, Stockholm, Sweden
Title: New methylation assay LUMA and its clinical applications

Steve Rapko
Staff Scientist
Genzyme Biosurgery, Cambridge, MA, USA
Title: DNA methylation study of chondrocytes and synovial fibroblast cultures

Shuji Ogino, MD., PhD.
Associate Pathologist & Assistant Professor of Pathology
Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA
Title: Epigenomic profiling of colorectal cancer in a large scale study

Manel Esteller, M.D., Ph.D.
Director, Cancer Epigenetics Laboratory,
Spanish National Cancer Centre (CNIO), Madrid, Spain
Title: The Epigenomes of Cancer Cells

Mukesh Verma, PhD.
Acting Chief of Analytic Epidemiology Research Branch &
Program Director, Epidemiology and Genetics Research Program
National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
Title: Cancer epidemiology and epigenetics

Christopher Adams, PhD.
Scientist, Epigenetics Division
Invitrogen Corporation, Carlsbad, CA, USA
Title:Title: siRNA targeting of exons results in epigenetic modifications of the chromatin at the corresponding gene

Marc Bühler, PhD.
Fellow in the Department of Cell Biology
Harvard Medical School, Boston, MA, USA
Title: Heterochromatic gene silencing

Kevin V. Morris, Ph.D.
Assistant Professor
Division of Rheumatology, The Scripps Research Institute, La Jolla, CA, USA
Title: A low-copy promoter RNA is required for siRNA directed transcriptional gene silencing in Human cells

Bradley E. Bernstein, M.D. Ph.D.
Assistant Professor of Pathology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA
Title: Chromatin state maps of pluripotent and lineage-committed cells

Alex Meissner, Ph.D.
Postdoctoral Fellow from the Lab of Prof. Rudolf Jaenisch
Whitehead Institute for Biomedical Research
Massachusetts Institute of Technology, Cambridge, MA, USA

Laurie Jackson-Grusby, Ph.D.
Assistant Professor Pathology
Children's Hospital Boston, Harvard Medical School, Boston, MA, USA

Jeffrey M. Besterman, Ph.D.
Executive Vice President & Chief Scientific Officer
MethylGene Inc., Canada
Title:MGCD0103, A Novel HDAC Inhibitor: From Bench to Clinic

Rene Cortese
Epigenomics AG, Berlin, Germany
Title: Conservation of tissue-specific DNA methylation patterns in human/mouse orthologues

Amit Meller, PhD.
Associate Professor of Physics and Biomedical Engineering
Boston University, Boston, MA, USA
Title: Progress toward ultra fast DNA sequencing using Nanopore arrays

Matthew Poulin, PhD.
Scientist
EpigenDx, Inc., Worcester, MA 01606 USA
Title: Pyrosequencing® Methylation Analysis of Multiple Genes on Limiting Amounts of DNA from Clinical Tissues and Cell Lines

Thomas L. Fare, PhD.
Director, Advanced Technologies Solutions
Rosetta Inpharmatics LLC., Merck, Seattle, WA 98109, USA
Title: Methods to evaluate DNA methylation profiling and transcription-factor binding on DNA microarrays

Jeffrey Falk, PhD.
Director of Technology & Business Applications
Aviva Systems Biology, San Diego, CA, USA
Title:  Profiling Epigenetic Modifications using ChIP-DSL, A Technology Enabling Genome-wide Characterization of Promoter Interactions and Modifications 

François Gaudet, Ph.D.
Research Investigator in Epigenetics, Novartis Institutes for Biomedical Research
Cambridge, MA, USA
Title: TBA

Menzo Havenga, Ph.D.
Vice President Research
Crucell Holland BV, Leiden, The Netherlands
Title: STAR and STAR-select technology to increase yield and expression stability of proteins in mammalian cells

Gerald Schock, Ph.D.
Global Product Manager Epigenetics
QIAGEN GmbH, Hilden, Germany
Title: Improved methylation analysis through prevention of DNA fragmentation during cytosine conversion

Mathias Ehrich, M.D., Ph.D.
Sequenom, Inc., San Diego, CA, USA
Title: Epigenetic Marker Discovery and Validation

Marina Bibikova, Ph.D.
Staff Scientist
Illumina, Inc. San Diego, CA, USA
Title: DNA Methylation Profiling of 1536 CpG Sites Across 96 Samples Using Illumina’s GoldenGate® Methylation Platform

Arturas Petronis MD., PhD
Associate Professor & Head
The Krembil Family Epigenetics Laboratory
University of Toronto-Centre for Addiction and Mental Health
Toronto, Ontario, Canada
Title: TBA

Jeffrey A. Jeddeloh, PhD.
Director, Science and Technology
Orion Genomics, St. Louis, MO, USA
Title: Massively parallel bisulphate pyrosequencing reveals the molecular complexity of breast cancer associated cytosine methylation patterns obtained from tissues and serum DNA

Alon M. Goren, MSc. (PhD).
PhD. Student in Professor Howard Cedar’s lab
Department of Cellular Biochemistry, Hebrew University Medical School
Jerusalem, 91120, Israel
Title: DNA replication timing of the human beta globin domain is controlled by histone modification at the origin

Benjamin G. Schroeder, PhD.
Senior Staff Scientist
Applied Biosystems, Inc.
Foster City, CA 94404, USA
Title: Advances in CE based methylation analysis

Julia Schliwka PhD.
Scientific Marketing Manager
Febit biotech GmbH, Heidelberg, Germany
Title: TBA

And many more from Biotech and Large pharma…..

Each speaker will have 25 min for presentation and 5 min for discussion.

Panel Discussion with experts from:
- Venture Capital Firm
- Technology Transfer Office
- Professional Science/Business Journalists
- Patent Attorney from a Law Firm
and selected speakers from the conference.

Exhibitors are welcome to reserve their booth space early!

GeneExpression Systems, Inc.
P.O. Box 540170
Waltham, MA 02454 USA
Tel: (781) 891-8181
Fax: (781) 891-8234
E-mail: Genexpsys@expressgenes.com
www.expressgenes.com

Poster Abstract Submission by June 22, 2007

All Abstracts

Global DNA-methylation analysis in vitro and in disease using LUMA
Tomas J Ekström, Ph.D. Professor, Department of Clinical Neuroscience, Karolinska Institutet, S-171 76 Stockholm, Sweden

Epigenetic states guide the interpretation of the genome. The heritable modifications of DNA and DNA-associated proteins involved, are endpoints of intracellular signaling, and are thus the link between the genome and the surrounding environment, intra- as well as extra-cellular. DNA-methylation is an important part of the epigenetic pattern and can be used to monitor physiological and pathological states. We have used our newly developed technique, LUMA, to show that global DNA-methylation can be used for mortality prognosis in chronic kidney disease. Investigations in vitro also show that inhibition of histone deacetylation causes rapid global and specific demethylation suggesting selective processes.

DNA methylation study of chondrocytes and synovial fibroblast cultures
Steve Rapko, Staff Scientist II, Genzyme Biosurgery, Cambridge, MA 02139, USA

Steve Rapko1, Udo Baron2, Uli Hoffmueller2, Leslie Wolfe1, and Sven Olek2
1Genzyme Biosurgery, USA; 2Epiontis GmbH, Germany

In this work, we analyzed DNA from forty seven strains of chondrocytes and eighteen strains of synoviocytes at various passages in monolayer culture, as well as DNA derived from cartilage tissue. Using bisulfite sequencing in conjunction with a statistically derived classifier system analyzing DNA methylation over seven genomic regions, all of the samples were grouped according to lineage with a high level of discrimination between cell types. This study demonstrates the powerful cell lineage identifying ability of DNA methylation analysis.

Epigenomic profiling of colorectal cancer in a large scale study
Shuji Ogino, MD., PhD. Associate Pathologist at the Department of Pathology, Brigham and Women's Hospital, Assistant Professor of Pathology, Harvard Medical School
Boston, MA, USA

CpG island methylator phenotype (CIMP) has been established as a distinct phenotype in colorectal cancer. To discriminate biologically significant methylation from insignificant (low level) methylation, quantitative assays are necessary. Methylation-specific real-time TaqMan PCR (MethyLight) are robust with good precision, and useful for paraffin-embedded tumor tissue in large scale studies. Discussion includes pros and cons for methylation-specific PCR against methylation-independent PCR. Quantitative PCR on carefully-selected CpG islands enables us to precisely diagnose CIMP status in a cost-effective way, and can be utilized to assess effects of CIMP on clinical outcomes – patient survival and treatment efficacy.

siRNA targeting of exons results in epigenetic modifications of the chromatin at the corresponding gene
Christopher Adams, PhD. Division of Epigenetics, Invitrogen Corporation, Carlsbad, CA, USA

Sharon Santos1, Mason Brooks1, Kevin V. Morris2, and Christopher Adams1
1Division of Epigenetics, Invitrogen Corporation, Carlsbad, CA, USA
2Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, USA
Small interfering RNAs (siRNAs) targeted to promoter regions in human cells can mediate transcriptional gene silencing (TGS) by directing histone modifications characteristic of silent chromatin. The extent to which siRNAs targeted to exonic regions can induce epigenetic modifications has remained unknown. Here we describe the transcriptional and post-transcriptional (PTGS) gene silencing of the Ubiquitin ligase promoter (UBC-1) in Human cells. SiRNA targeting of both the promoter and exon 1 of UBC-1 resulted in gene silencing that correlated with H3 lysine-9 di-methylation (H3K9me2+) and H3 lysine-27 tri-methylation (H3K27me3+) at the siRNA-targeted UBC-1 promoter and suprisingly also at the UBC-1 exon. The addition of Trichostatin A resulted in an abrogation of siRNA mediated TGS of the UBC-1 promoter but had little effect on siRNA mediated PTGS of the UBC-1 exon-1. These data suggest that siRNA targeting of mRNAs in a PTGS manner also produce an epigenetic off-target effect at the local chromatin of the corresponding gene and that there are distinct mechanistic differences between TGS and PTGS in human cells.

Conservation of tissue-specific DNA methylation patterns in human/mouse orthologues.
Rene Cortese, Epigenomics AG, Berlin, Germany

DNA methylation represents the most stable epigenetic modification modulating the transcriptional plasticity of mammalian genomes. We have determined DNA methylation profiles of human chromosomes 6, 20 and 22 in 12 different healthy tissues and primary cells by direct bisulfite sequencing. We have found 470 tissue-specific, differentially methylated regions (T-DMRs) (p<0.001) located within 5’unstranslated regions (5’-UTR), exons, introns and non-genic, evolutionarily conserved regions (ECRs). As human chromosome 6 is highly syntenic to mouse chromosome 17, we investigated the conservation of 59 TDMRs corresponding to 5’-UTRs and non-genic ECRs in the corresponding murine orthologue. The majority (70%) of the amplicons differed by less than 20% methylation, indicating significant conservation. Our findings add further evidence to the concept that epigenetic states may be evolutionarily conserved among mammals.

Heterochromatic gene silencing
Marc Bühler, PhD. Fellow of Cell Biology, Harvard Medical School, Department of Cell Biology, Boston, MA, USA

Heterochromatin is an altered chromatin structure that is epigenetically inherited and plays important roles in gene regulation and genome stability. The long standing paradigm that heterochromatin is transcriptionally inert has been challenged by recent studies suggesting that RNA interference (RNAi)-dependent gene silencing in heterochromatin can be mediated primarily by degradation of nascent transcripts within these domains. Data will be presented that demonstrate that transgenes inserted in heterochromatic repeats are direct targets of RNAi. In addition, a role for another RNA degradation pathway involving the TRAMP and exosome complexes in robust silencing of heterochromatic genes will also be described.

A low-copy promoter RNA is required for siRNA directed transcriptional gene silencing in Human cells
Kevin V. Morris, PhD. Assistant Professor, Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, USA

Small interfering RNAs (siRNAs) targeted to gene promoters can direct epigenetic modifications that result in transcriptional gene silencing in human cells. It is not clear whether the antisense strand of the siRNAs binds DNA or to an RNA transcript corresponding to the promoter region. We present direct evidence that a species of previously uncharacterized low-copy RNAPII expressed RNAs is required for siRNA directed epigenetic modifications and transcriptional silencing of siRNA targeted promoters. These data demonstrate the target moiety for RNA directed control of gene expression and suggest an RNA component is involved in writing the histone code.

Cancer Epidemiology and Epigenetics
Mukesh Verma, Ph.D., Acting Chief of Analytic Epidemiology Research Branch, Program Director. Epidemiology and Genetics Research Program, Division of Cancer Control and Population Sciences, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

Cancer is a genetic and epigenetic. Identification of populations at risk of developing cancer is important as it provides opportunities for prevention and treatment of cancer. Gene expression changes during the initiation, progression, and development of cancer. To identify different stages of tumor development markers are utilized. The most common markers belong to two categories: genetic and epigenetic. Genetic changes arise due to change in nucleotide sequence whereas epigenetic changes occur due to changes in chromatin conformation and methylation the CpG islands located primarily in the promoter region of a gene. Epigenetic markers exhibit high sensitivity and specificity for different tumor types and can be assayed in biofluids and other specimens collected by non-invasive technologies. In colon and breast cancer epigenetic markers have been utilized successfully to identify high risk populations. Research opportunities in epigenetics field at the National Cancer Institute will be discussed.

Profiling Epigenetic Modifications using ChIP-DSL, A Technology Enabling Genome-wide Characterization of Promoter Interactions and Modifications 
Jeffrey Falk, PhD., Director of Technology & Business Applications, Aviva Systems Biology, San Diego, CA, USA

We have developed a novel promoter array technology, ChIP-DSL (Chromatin Immunoprecipitation – DNA Selection and Ligation), that facilitates genome-wide profiling of Epigenetic Modifications, DNA Methylation Sites and transcription factor/promoter interactions involved in important systems and pathways. These studies used global promoter mapping and locus tiling arrays to offer new insight into transcriptional and epigenetic mechanisms involved in cancer, and provide a powerful tool with which to decode the molecular underpinnings of diseases such as cancer, as well as stem cell differentiation.

Pyrosequencing® Methylation Analysis of Multiple Genes on Limiting Amounts of DNA from Clinical Tissues and Cell Lines
Matthew Poulin, Ph.D., Scientist, EpigenDx, Inc., Worcester, MA 01606 USA

Because clinical tissues samples are treated in different ways and the quantity is often limiting, it is important to show that consistent methylation results can be obtained with small quantities of DNA. In this study we quantitatively analyzed both global and gene specific methylation of DNA from different tissue preparations, as well as a variety of cell lines. Among the genes we analyzed were the Alu and Line-1 elements for global methylation and gene specific MGMT, p16 and RASSF1 assays. We demonstrated that Pyrosequencing® methylation analysis achieves a high degree of consistency and sensitivity when used on DNA isolated from clinical tissues and different cell lines.

Comprehensive gene regulation studies using 454 Sequencing™
Maithreyan Srinivasan, PhD. Director, Amplification and Sequencing Technology, 454 Life Sciences-Roche Diagnostics, Branford, CT USA

454 Sequencing™ (pyrophosphate-based sequencing optimized for beads in picoliter wells) involves subjecting template DNA to solid-phase amplification on to beads in emulsions. Beads that contain amplified template DNA are deposited into the wells of a picotiter plate (fused fiber-optic reaction vessels arranged in tandem) and 454 Sequencing™ is performed to obtain reads with average read lengths of 100 or 250-bases. Data obtained from investigations that quantify methylation patterns, detect rare drug resistant HIV mutants in patient samples, discover new miRNAs and new classes of small RNAs, as well as shed light on the first individual human genome will be presented.

Massively parallel bisulphite pyrosequencing reveals the molecular complexity of breast cancer associated cytosine methylation patterns obtained from tissue and serum DNA
Jeffrey.A. Jeddeloh, PhD. Director of Science and Technology, Orion Genomics, LLC., St. Louis. MO, USA

Y. Korshunova, R.K. Maloney, N. Lakey, R.W. Citek, B. Bacher, A. Budiman, J.M. Ordway, W.R. McCombie, J. Leon, J.A. Jeddeloh* and J.D. McPherson1
Orion Genomics, LLC., St. Louis. MO, USA
1 Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA

Cytosine-methylation changes are stable and thought to be among the earliest events in tumorigenesis. Theoretically, DNAs carrying tumor-specifying methylation patterns escape the tumors and may be found circulating in the sera from cancer patients; thus, providing the basis for development of non-invasive clinical tests for early cancer detection. Indeed, using PCR-based techniques, several groups reported the detection of tumor-specifying methylated DNA in the sera from cancer patients with varying clinical success. However, by design such analytical approaches allow assessment of the presence of molecules with only one methylation pattern, leaving the bigger picture unexplored. The limited knowledge about circulating DNA methylation patterns hinders the efficient development of clinical methylation tests and testing platforms. Here, we report the results of a comprehensive methylation pattern analysis from breast cancer clinical tissues and sera obtained using massively-parallel bisulphite-genomic pyrosequencing. The four loci studied were recently discovered by our group, and demonstrated to be powerful epigenetic biomarkers of breast cancer. The detailed analysis of more than 700,000 DNA fragments derived from more than 50 individuals (with cancer as well as cancer-free) revealed an unappreciated complexity of genomic cytosine methylation patterns in both tissue derived and circulating DNA, and has better defined the development challenges facing DNA methylation based cancer-detection tests.

DNA replication timing of the human beta globin domain is controlled by histone modification at the origin
Alon M. Goren, MSc. (PhD). Student in Professor Howard Cedar’s lab, Department of Cellular Biochemistry, Hebrew University Medical School, Jerusalem, Israel

Alon Goren, Amaliya Tabib, Merav Hecht, and Howard Cedar
Department of Cellular Biochemistry, Hebrew University Medical School, Jerusalem, Israel

The human beta globin genes constitute a large chromosomal domain that is developmentally regulated. In non-erythroid cells these genes are packaged in an inaccessible chromatin structure and replicate late in S-phase, while in erythroid cells, the entire domain replicates early. ChIP analysis shows that the replication origin region located near the b gene is differentially packaged with acetylated histones in erythroid cells, yet is associated with deacetylated histones in non-erythroid cells. Recruitment of a histone acetylase (HAT) to this origin in vivo brings about chromatin reacetylation in fibroblast or lymphoid cells, and this is accompanied by a shift to early replication. In contrast, tethering of a histone deacetylase (HDAC) to this same origin region in erythroblasts causes a decrease in the level of histone acetylation and a concomitant shift to late replication. These results suggest that histone modification at the origin serves as a switch for controlling replication timing.

DNA Methylation Profiling of 1536 CpG Sites Across 96 Samples Using Illumina’s GoldenGate® Methylation Platform
Marina Bibikova, Ph.D. Staff Scientist, Illumina, Inc., San Diego, CA, USA

DNA methylation is widespread and plays a critical role in the regulation of gene expression in cancer, development, differentiation and various diseases. The ability to access the epigenomic information for a large number of genes or the entire genome should greatly facilitate the understanding of the nature of gene regulation in cells, and epigenomic mechanism of interactions between cells and environment. We developed a DNA methylation detection method based on “genotyping” of bisulfite-converted genomic DNA. In this assay, non-methylated cytosines (C) are converted to uracil (U) when treated with bisulfite, while methylated cytosines remain unchanged. The detection of the methylation status of a particular cytosine in the genome can be carried out using a genotyping assay for a C/U polymorphism. The GoldenGate Assay for Methylation combines a high level of assay multiplexing and scalable automation for sample handling and data processing with a miniaturized bead-based array platform. The throughput of the system is 96 samples per run in a microtiter plate format. Several plates can be processed in parallel. The assay sensitivity and specificity is sufficient to analyze methylation status of up to 1536 CpG loci simultaneously with 250 nanograms of human genomic DNA. Reproducible DNA methylation profiles were obtained within replicates (an average r2 of 0.98).

Two papers have been published in Genome Research using this assay. We designed assays for 1536 CpG sites from the 5'-regulatory regions of 371 genes selected for biological relevance to investigate specific methylation patterns in cancer cell lines and to identify methylation markers in lung adenocarcinomas. We have also assessed the epigenetic signature of human embryonic stem cells and identified a specific set of CpG markers to monitor hES cell differentiation. In January, Illumina commercially launched a standard Methylation Cancer Panel I to interrogate 1505 CpG sites selected from 807 genes. These genes fall into various classes, including tumor-suppression genes, oncogenes, genes involved in DNA repair, cell cycle control, differentiation, apoptosis, X-linked, and imprinted genes. In collaboration with the University of Kiel (Germany), we used this panel to study DNA methylation changes associated with classical Hodgkin lymphoma (cHL) in comparison with normal hematological tissues. Methylation profiling revealed a global alteration of the methylome in HL cell lines, being mostly characterized by promoter hypermethylation of a large number of genes. Hypermethylated gene promoters in HL included known tumor suppressor genes (e.g. p16, p73, DAPK). Interestingly, CpG-islands from genes involved in B-cell specific pathways (e.g. BCAM, BLK, MME and SYK) were hypermethylated in cHL. Such epigenetic silencing of B-cell specific genes may be the cause of loss of the B-cell identity characteristic for cHL, and thus, might play a key role in its pathogenesis. The methylation data generated by the array was validated by methylation specific PCRs and bisulfite sequencing.

The GoldenGate Assay for Methylation will provide powerful insight into epigenetic mechanisms of gene regulation that can be applied to diagnosis, prognosis, and treatment of diseases.

Improved methylation analysis through prevention of DNA fragmentation during cytosine conversion
Gerald Schock, PhD. Global Product Manager Epigenetics, QIAGEN GmbH, Germany

Accessing epigenetic information is of prime importance for many areas of biological and medical research — particularly oncology but also stem cell research and developmental biology. However, the analysis of changes in DNA methylation is challenging due to the lack of standardized methods for providing reproducible data from limited sample material (e.g., patient biopsies or FFPE material). This presentation will provide background information on standardized workflows in epigenetic analysis and will highlight steps of critical importance — such as the complete conversion of unmethylated cytosine to uracil that can be readily detected by sequencing or PCR — for reproducible and accurate results.