Zymo-Seq Cell Free DNA WGBS Library Kit
24 Preps
Zymo D5462

Zymo-Seq Cell Free DNA WGBS Library Kit
24 Preps
Zymo D5462

Zymo-Seq Cell Free DNA WGBS Library Kit provides an optimized and reliable workflow for whole genome bisulfite sequencing (WGBS) library preparation from cell-free DNA (cfDNA). This all-inclusive kit features a straightforward procedure capable of preparing high-quality methyl-seq libraries from as little as 5 ng of cfDNA. The process is completed in three basic steps: bisulfite conversion, direct adapter ligation, and index PCR amplification. The initial bisulfite treatment is gentle on cfDNA yet effective in converting any unmodified cytosines into uracil. Next, the innovative splinted adapters capture and directly ligate the Illumina-compatible adapters onto any size DNA fragment, allowing for sequencing of nicked, damaged, and short DNA fragments that may have otherwise been discarded with traditional library prep methods. The direct ligation of the adapters also eliminates the need for second strand synthesis, end repair, and dA tailing steps, thus reducing bias by preserving the integrity of any native methylation that is present on the fragment termini. Finally, the adapter-ligated cfDNA is indexed and amplified via PCR with Zymo-Seq UDIs, generating libraries that are ready for sequencing on any Illumina instrument.

Optimized for small fragment input: Ideal for small and damaged DNA fragments such as cell-free DNA (cfDNA). Accurate methylation calling: Direct ligation-based protocol allows for accurate reads and methylation calling of native termini for each DNA fragment. Streamlined and simple workflow: Prepare robust methyl-seq libraries in as little as 3 steps.

Barcode Sequences Please refer to the Documents Section. Bisulfite Conversion >99.5% of non-methylated cytosine residues are converted to uracil; >99.5% protection of methylated cytosines. Equipment Required Thermal cycler(s) with temperature adjustable lids, microcentrifuge, magnetic stand. Hands-On Time ~2 hours Input Quality For optimal results, use at least minimum input of purified cfDNA with no RNA or gDNA contamination. cfDNA can be concentrated using the DNA Clean & Concentrator (D4013) prior to processing. Input cfDNA can be suspended in water, DNA Elution Buffer, or TE buffer. Library Storage Libraries eluted in DNA Elution Buffer (provided) may be stored at ≤ 4°C overnight or ≤ -20°C for long-term storage. Maximum Input 10 ng Minimum Input 5 ng Sample Input Material Purified cell-free DNA (cfDNA) Sequencing Compatibility Libraries are compatible with all Illumina sequencing platforms. Recommended: NextSeq, NovaSeq. Total Processing Time ~6 hours