Complete methylome analysis from only 50 ng of genomic DNA

Cambio, the Cambridge, UK-based supplier of world-class molecular biology

reagents and consumables, has launched the EpiGnomeTM Methyl-Seq Kit in the UK.

EpiGnome, developed by US biotech firm Epicentre, an Illumina company,

produces whole genome bisulfite sequencing libraries from as little as 50 ng

of genomic DNA. The kit uses a unique 'post-bisulfite conversion' library

construction method that yields high diversity libraries with uniform

coverage.

The process of bisulfite treatment denatures genomic DNA (gDNA) into

single-stranded DNA (ssDNA). EpiGnome converts all the ssDNA into an

IlluminaR sequencing library, eliminating the sample loss associated with

other methods.

The elimination of sample loss using EpiGnome means that as little as 50 ng

of total DNA is needed to detect methylation. Uniform coverage is clearly

seen at this scale (Figure 1).

EpiGnome has the sensitivity to detect methylation changes with only 50 ng

of input gDNA. When 50 ng of lymphoblastoid gDNA was treated with bisulfite

and converted into a sequencing library using EpiGnome (first example), the

CpG methylation pattern across regions of chromosome 1 highlighted areas of high CpG methylation (red) and low CpG methylation (blue). Similarly, when 50 ng of a hypermethylated HeLa sample was treated with bisulfite and

converted into a sequencing library using EpiGnome (second example), most

reads showed methylation (red), and very few reads were not methylated

(blue).

EpiGnome offers uniform CpG, CHG and CHH coverage. The method does not require fragmentation by Covaris ultrasonication nor methylated adaptors, as required by other bisulfite sequencing library prep methods. In addition to

eliminating sample loss, the EpiGnome method takes only 5 hours, a fraction

of the time taken by alternative methods.