What is the correct order of addition of Infinium Amplification reagents?

The Infinium assay workflow includes a critical DNA amplification step. Before amplification, genomic DNA must be combined with MA1 and NaOH to facilitate denaturation and stabilization of the reaction. MA1 plays a key role in preventing evaporation of both NaOH and DNA during incubation and thus,must always be added first, regardless of assay type.

Recommended order of adding amplification reagents:

• Infinium HD and HTS Assays: MA1 > DNA > NaOH > MA2 > MSM

• Infinium HD Methylation and LCG Assays: MA1 > DNA > NaOH > RPM > MSM

• Infinium XT Assay : MA1 > NaOH > DNA >MA2 > RAM

• Infinium EX Assay: MA1 > NaOH > DNA > MA2 > AAX

• Infinium EX Methylation Assay: MA1 > NaOH > DNA >MPM > MSM

These reagent addition sequences have been validated by Illumina. Adhering to the correct order of reagent addition ensures consistent and high-quality assay performance. Illumina does not guarantee assay performance if the protocol is not followed as specified as deviations may negatively impact results.

Refer to the Infinium Product Documentation for more information:

• Infinium HD Assay Ultra Protocol Guide

• Infinium HD Super Assay Reference Guide

• Infinium HD Methylation Assay Product Documentation

• Infinium HTS Assay Reference Guide

• Infinium HTS Extra Reference Guide

• Infinium LCG Assay Product Documentation

• Infinium LCG Quad Assay Protocol Guide

• Infinium XT Assay Reference Guide: ST Workflow | HT Workflow

• Infinium EX Assay Product Documentation

• Infinium EX Methylation Assay Product Documentation

For any feedback or questions regarding this article (Illumina Knowledge Article #5874), contact Illumina Technical Support [e-mail].