DNA Methylation Profiling Using Infinium Methylation Assay

Materials and Reagents

1. EZ-96 DNA Methylation Kit (Zymo Research, catalog number: D5003 )
   
2. Ethanol (VWR International, catalog number: 20821.330 )
   
3. Genomic DNA
   
4. PCR Grade water (Life Technologies, Invitrogen^TM, catalog number: AM9937 )
   
5. PCR reagents: PCR buffer, dNTPs, Taq (Roche Applied Science, catalog number: 04738420001 )
6. Primers for methylation specific PCR [see Step 20-(1)]
   
7. 96-well 0.2 ml microplates (Starlab, catalog number: I1402-9800-C )
   
8. 96-well 0.8 ml microplates (Thermo Fisher Scientific, catalog number: AB-0765 )
   
9. Seal sheets (Thermo Fisher Scientific, catalog number: AB-0558 )
   
10. Infinium Human Methylation450 BeadChip Kit (Illumina, catalog number: WG-314-1003 )
    
11. PyroMark Q96 CpG LINE-1 Kit (QIAGEN, catalog number: 973043 ) and associated reagents (Optional)
    

Equipment

1. 96-well Thermal Cycler
   
2. Centrifuge with microplate adapters
   
3. Vortex mixer
   
4. HiScan SQ or iScan systems (Illumina, catalog number: SY-103-2001 or SY-101-1001 respectively)
   
5. GenomeStudio Methylation Module software (Illumina, catalog number: SW-300-1001 )
   
6. NanoDrop ND-1000 UV-Vis spectrophotometer (Thermo Fisher Scientific) or equivalent
   
7. Speed Vacuum (Optional)
   
8. PyroMark Q96 MD (QIAGEN, catalog number: 9001526 ) or equivalent (Optional)
   

Procedure