Real-time PCR-based genotyping from whole blood using Taq DNA polymerase and a buffer supplemented with 1,2-propanediol and trehalose
Pavol Utekal, Lukas Kocanda, Petr Matousek, Petr Wagner, Petr Draber, Viktor Bugajev
DOI: 10.1016/j.jim.2014.09.006
Klíčová slova: 1,2-Propanediol; Real-time PCR; SYBR green I; Taq DNA polymerase; Trehalose; Unseparated blood
Anotace: Amplification of DNA templates from whole blood with Taq DNA polymerase remains a difficult task worldwide. Using a real-time PCR setup and a buffer supplemented with 1M 1,2-propanediol, 0.2M trehalose, and SYBR green I we show a reliable technique for genotyping in mice and detection of single-nucleotide polymorphisms/mutations in humans. Elimination of DNA extraction and use of the common Taq DNA polymerase and DNA dye bring about substantial savings in labor and cost.
Citace: UTEKAL, Pavol, Lukas KOCANDA, Petr MATOUSEK, Petr WAGNER, Petr DRABER a Viktor BUGAJEV. Real-time PCR-based genotyping from whole blood using Taq DNA polymerase and a buffer supplemented with 1,2-propanediol and trehalose. Journal of Immunological Methods. Amsterdam: Elsevier Science Publishers, 2015, 416(-), 178-182. ISSN 0022-1759. Dostupné z: doi:10.1016/j.jim.2014.09.006