Epigenetic Monitoring
Epigenetic factors are known to play a critical role in regulating stem cell differentiation by controlling when and how transcription factor bind to promoters or other regulatory elements. Understanding this regulatory mechanism will provide insight into how cells, containing the same genetic material, exhibit significantly different characteristics.
Genetic modifications include the following:
- Genomic imprinting
- X-chromosome inactivation
- DNA methylation
- Histone modifications
These modifications possess unique molecular signatures that are characteristic by cell state (self-renewal vs. differentiated) or cell type (embryonic, early progenitors or lineage specific cells).
DNA Methylation Analysis is used to identify patterns of methylation by relying on bisulfite treatment of methylated genomic DNA to convert unmethylated cytosine to uracil. Methylated cystosine residues remain unchanged. Treated or untreated samples can be directly compared to identify methylated sites and unique patterns of methylation within a site.
The Applied Biosystems methylSEQr™ Bisulfite Conversion Kit Workflow is based on PCR amplification and direct sequencing to offer greater resolution to identify unique molecular signatures. The methylSEQr™ kit protocol consists of the following:
Experimental Workflow: Find the Products You Need for Every Step
Click the arrows above to view products
Step 1: Design Primer
Design high quality PCR primers for methylation mapping experiments using Methyl Primer Express® Software v1.0. Simply cut and paste in your region of interest. The tool searches for CpG islands and simulates bisulfite modification of DNA in silico. Applied Biosystems supplies all the tools you need to complete methylation mapping experiments.
Products for Step 1: Design Primer
-
Methyl Primer Express® Software v1.0
-
This free software enables you to design high quality PCR primers for methylation mapping experiments. Simply cut and paste in your region of interest. The tool searches for CpG islands and simulates bisulfite modification of DNA in silico. Applied Biosystems supplies all the tools you need to complete methylation mapping experiments.
Step 2: Recover DNA
The methylSEQr™ Bisulfite Conversion Kit converts non-methylated cytosines (C) in your DNA sample to uracil (U). This kit is an initial step in a multi-step workflow that enables you to determine which cytosines in your sample are methylated by comparing the sequences of treated vs. non-treated DNA.
Products for Step 2: Recover DNA
-
methylSEQr™ Bisulfite Conversion Kit
-
The methylSEQr™ Bisulfite Conversion Kit converts non-methylated cytosines (C) in your DNA sample to uracil (U). This kit is an initial step in a multi-step workflow that enables you to determine which cytosines in your sample are methylated by comparing the sequences of treated vs. non-treated DNA.
Step 3: Amplify
Identify Unique Molecular Signatures using a Streamlined workflow based on BigDye® Terminator v1.1/v3.1 Sequencing Chemistry, BigDye® XTerminator™ Purification Kits, Applied Biosystems Thermal Cyclers and Genetic Analyzers.
Products for Step 3: Amplify
-
GeneScan™ 500 LIZ® Size Standard
-
The GeneScan™ 500 LIZ® Size Standard is a fifth dye-labeled size standard for the reproducible sizing of fragment analysis data. Use this size standard for fragments between 35 and 500 bp. The standard contains 16 LIZ® dye-labeled, single-stranded DNA fragments. Since the standard is labeled with the fifth dye, users can genotype a greater number of markers in a given lane, compared to the four-dye system.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
on invitrogen.com 
