Synthetic positive control for human KRAS gene

Synthetic positive control has emerged as the essential tool for modern qPCR testing kit as a means for the clinicians to verify that the test performs correctly. With declining cost of gene synthesis, more clinical diagnostic companies are adopting synthetic gene for developing their positive controls. In the case of human KRAS testing, the entire KRAS gene has been found to be too complex for synthesis by commercial DNA synthesis companies. This work provides a sequence fragment of the human KRAS gene that can be synthesized through commercial DNA gene synthesis. Primers could be designed for this gene fragment, and SYBR green dye chemistry could be used to detect amplicons from this template gene fragment with a dynamic range from 109 copies per µl to 10 copies per µl. The limit of detection is 10 copies per µl with a Ct value of 40.