The ability of human embryonic and induced pluripotent stem cells to differentiate into all three germ layers has enormous potential for basic human developmental research and regenerative medicine. ScienCell has created a convenient qPCR kit for the assessment of human pluripotent stem cell differentiation. NEUROD1, ACTA2 and AFP qPCR primers included in the kit allow for the detection and quantification of ectoderm, mesoderm and endoderm, respectively1.Note: all gene names follow their official symbols by the Human Genome Organization Gene Nomenclature Committee (HGNC).Four qPCR controls are included in this kit to verify successful reverse transcription of messenger RNA (mRNA) to complementary DNA (cDNA), reveal the presence of genomic DNA (gDNA) contamination in cDNA samples, and detect qPCR inhibitor contamination. Good quality cDNA is a critical component for successful gene expression analysis.Each primer set included in HGL-qPCR kit arrives lyophilized in a 2 mL vial. All primers are designed and tested under the same parameters: (i) an optimal annealing temperature of 65°C (with 2 mM Mg2+, and no DMSO); (ii) recognition of all known target gene transcript variants; and (iii) specific amplification of only one amplicon. Each primer set has been validated by qPCR by melt curve analysis and gel electrophoresis.