Imprint® Methylated DNA Quantification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Ensure DNA is mixed thoroughly prior to application. The wells of the assay strip bind up to 200 ng of total DNA.

Protocol tips
Ensure DNA is mixed thoroughly prior to application. The wells of the assay strip bind up to 200 ng of total DNA.

Publication protocol

RRBS libraries were prepared from isolated DNA following published protocols [36]. Briefly, RRBS libraries were prepared by MspI digestion of 100–500 ng genomic DNA, followed by end-repair and T-tailing using Klenow Exo- (Fermentas). Adapter ligation was performed overnight (in-house adapters) using T4 DNA Ligase (NEB), followed by a clean-up step using AMPure XP beads (Agencourt, 0.9×). Subsequently, libraries were bisulfite-treated according to the manufacturer’s instructions (Sigma Imprint Kit; 2 step protocol) and purified using an automated liquid handling robotic system (Agilent Bravo). The libraries were amplified using KAPA HiFi Uracil HotStart DNA Polymerase (KAPA Biosystems), indexing the samples with individual primers. All amplified libraries were purified (AMPure XP beads, 0.8×) and assessed for quality and quantity using High-Sensitivity DNA chips on the Agilent Bioanalyzer. High-throughput sequencing of all libraries was carried out with a 75 bp paired-end protocol on a HiSeq 2000 instrument (Illumina).

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Imprint® Methylated DNA Quantification Kit below.

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