LINE-1 Methylation Analysis Alternative: MethylFlash™ Global DNA Methylation (5-mC) ELISA Easy Kit
Given the wide array of options to quantitate global DNA methylation, one method is Long Interspersed Nuclear Element-1 (LINE-1) methylation analysis. Here, the levels of methylation of LINE-1 repeats are determined often by bisulfite conversion, PCR amplification & pyrosequencing. Given the repetitive motifs of LINE-1 (comprising approximately 18-20% of the genome) the results are extrapolated to determine the global DNA methylation level.
EpiGentek is here to provide an alternative to make the process easier, quicker and more reliable. Our proprietary MethylFlash™ Global DNA Methylation (5-mC) ELISA Easy Kit provides everything you need to get quality data on global DNA methylation quantitation using an ELISA-like method. With a simple 2-hour procedure, our kit allows the flexibility for either manual or high throughput options. Simplified to 3 main steps in the protocol, you’ll be able to complete more research in less time.
Pyrosequencing and/or PCR amplification often required for analysis
No; assay provides complete set of buffers & reagents for experiment
DNA Denaturing Required?
Yes
No
DNA Methylation Analysis Basis
Based on methylation of LINE-1 repeats to extrapolate global DNA methylation status
Based on complete input DNA bound to assay strip-wells
Accurate quantification of 5-mC content of various DNA samples from different species using the MethylFlash™ Global DNA Methylation (5-mC) ELISA Easy Kit (Cat. #P-1030). The results are closely correlated with those obtained by HPLC-MS.
An example of an optimal standard curve generated with 5-mC standard control.
Full Advantages of the MethylFlash™ Global DNA Methylation (5-mC) ELISA Easy Kit
Fast - Reduced steps allow you to finish in only 2 hours
Robust - Improved kits have a greater “signal window” with reduced variation between replicates
Convenient - Inherently low background noise without the need for DNA denaturation and plate blocking steps
Sensitive - Detection limit can be as low as 0.05% methylated or 0.01% hydroxymethylated DNA from 100 ng of input DNA
Specific - No cross-reactivity to unmethylated or other modified cytosines
Universal - Included positive and negative controls allow for detection of DNA methylation or hydroxymethylation in any species from either single-stranded or double-stranded input DNA
Accurate - Optimized positive controls that can be fractionalized in percentage scale, allowing the assays to be more accurate and highly comparable with HPLC-MS analysis
Flexible - Strip-well microplate format for manual or high throughput analysis
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