Quantitative synthetic Hepatitis A virus DNA can be used for assay development, verification, and validation as well as monitoring of day-to-day test variation and lot-to-lot performance of molecular-based assays. The quantitative format allows for the generation of a standard curve for quantitative PCR (qPCR) to determine viral load. This preparation includes genomic segments from the 5’ untranslated region, viral capsid proteins (VP1- 4), self-cleaving peptide 2A, proteinase 3C, and 3D RNA polymerase.