Infinium EX Methylation DNA input guidelines and compatible bisulfite conversion kits

Supported Sample Types

• The Methylation Screening Array (MSA) is validated for blood samples.

• MSA is not validated with FFPE or saliva samples.

Recommended DNA Input

• The recommended DNA input for the bisulfite conversion reaction (manual) is minimum 50 ng, and up to 1 μg.

• For optimal results and higher reproducibility, use higher DNA input.

Refer to the Infinium EX Methylation Assay Product Documentation for additional information.

Supported Bisulfite Conversion Kits

Before carrying out the Infinium EX Methylation Assay, samples must first be bisulfite converted. Illumina has validated and recommends only the following kits from Zymo Research for Infinium EX Methylation.

EZ-96 DNA Methylation Lightning MagPrep Kits (uses magnetic beads for purification) Bisulfite conversion completed in ~ 3 hours

Product Name	Size	Format	Catalog No.	Elution Volume	Preparation
EZ-96 DNA Methylation-Lightning MagPrep	4 x 96 Rxns	96-well (Deep)	D5046	25 μl	Manual
EZ-96 DNA Methylation-Lightning MagPrep	8 x 96 Rxns	96-well (Deep)	D5047	25 μl	Manual

Note: Zymo EZ DNA Methylation kits (D5001, D5002, D5004) have not been validated and are not supported for use with Infinium EX Methylation.

Recommended DNA Sample Volumes for Zymo Bisulfite Conversion **For Zymo kits D5046 and D5047, the Zymo protocol states:**1. Add 130 µl of Lightning Conversion Reagent to 20 µl of a DNA sample in a Conversion Plate. Mix the samples by pipetting up and down. Note: If the volume of DNA is less than 20 µl, compensate with water.

Note: DNA quantification must be performed before bisulfite conversion. After bisulfite conversion, the DNA does not need to be quantified and the user can proceed directly to the Infinium EX Methylation Assay.

Additional information: * For information on how the Lightning kits were validated, refer to the Illumina Technical Note: Automated bisulfite conversion for Infinium Methylation BeadChips

For any feedback or questions regarding this article (Illumina Knowledge Article #8604), contact Illumina Technical Support [e-mail].