methylRAD-Seq
Powered by NEBNext® Enzymatic Methyl-seq Conversion Module
WHAT IS methylRAD-SEQ?
MethylRAD is an epigenetic modification to the traditional RAD-Seq protocol for assaying the methylation status of cytosines. Like RAD-Seq, the same subset of a genome is sequenced across a population of samples, but late in the library preparation all unmethylated cytosines are converted to thymines, allowing population-level assessment of differential methylation. Traditionally, the cytosine to thymine conversion is performed with sodium bisulfite, but Floragenex makes use of an enzymatic conversion module which causes less degradation and fragmentation of the treated DNA. Currently, a reference sequence in the form of a whole genome or prior RAD-Seq data is required to map the data and determine methylation status.
APPLICATIONS
Epigenetic studies where general methylation patterns are being studied. For example:
Population genomics
Evolution
Phenotypic plasticity and environmental adaptation
Cancer genomics in non-model systems
EXAMPLE methylRAD-SEQ PROJECT
Population Based Epigenomics
Surveyed >60 samples to look at epigenetic differences across multiple populations with color variation
Used SbfI to fragment the genome, to 20,000 loci; library is converted with NEB EM-Seq Conversion Module
Illumina Nova Seq 6000 2x150bp SP
Looking for differential methylation patterns acorss populations that are correlated with different color patterns