Human APOE Genotyping qPCR Assay Kit

Apolipoprotein E (ApoE) is a key protein involved in lipid transport and cholesterol homeostasis in both the brain and peripheral tissues. Variations in the APOE gene, resulting from two single nucleotide polymorphisms (rs429358 and rs7412), give rise to three major alleles (ε2, ε3, and ε4) that influence susceptibility to cardiovascular and neurodegenerative diseases. ScienCell’s Human APOE Genotyping qPCR Assay Kit (APOE) enables direct genotyping from cells without DNA isolation, using a nested PCR strategy followed by allele-specific amplification to determine the genotype. This design ensures high specificity and sensitivity, with positive and negative reactions fully concordant with sequencing results. The assay is robust, reliably genotyping samples with a limited number of cells.

The APOE kit can use either cell lysate or purified genomic DNA as the PCR template. The included cell lysis buffer and enhancer (Cat# GQ400a and GQ400b) are used to lyse pelleted cell samples. MB6038 master mix is provided for the initial PCR reaction, and the 2X GoldNStart TaqGreen qPCR Master Mix (Cat# MB6018a-1), a SYBR® Green–based master mix, is supplied for the qPCR step. The Nested PCR Outer Primer Set amplifies a larger region of the APOE and ACTB genes, while the E2, E3, and E4 primer sets detect the corresponding APOE alleles. The ACTB primer set amplifies the beta actin gene as an internal control. All primer sets have been validated by PCR and gel electrophoresis for amplification specificity. The reference positive control consists of plasmids containing the E2, E3, and E4 APOE alleles along with the ACTB gene, and serves as a positive control for the assay.