# 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](https://support.illumina.com/downloads/infinium-ex-methylation-assay-product-documentation.html) 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](http://www.zymoresearch.com/) 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](https://www.illumina.com/content/dam/illumina/gcs/assembled-assets/marketing-literature/automated-bisulfite-infinium-methylation-tech-note-m-gl-00144/automated-bisulfite-Infinium-methylation-tech-note-m-gl-00144.pdf)

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