Authors:Mitja Križman, Jernej Jakše, Mirko Prošek, Dea Baričevič, and Branka Javornik
Agarose gel electrophoresis is a basic separation tool used in molecular biology, mostly for qualitative DNA analysis. There are constraints limiting its use in quantitative analysis, namely low repeatability and a narrow linear range. However, by using an internal standard or internal normalization, repeatability and linear range could be significantly improved. In the work discussed in this paper it was shown that an approximately fivefold improvement in repeatability and an over threefold wider linear range could be achieved by applying internal normalization. Using the proposed approach, genetic markers, for example RAPD and PCR-RFLP, or even microsatellite markers, could be conveniently quantitatively assessed using agarose gel electrophoresis.
Authors:Katsuyuki Tokinoya, Seiko Ono, Kai Aoki, Koki Yanazawa, Yasuhiro Shishikura, Takehito Sugasawa, and Kazuhiro Takekoshi
for 3 s, and annealing and elongation at 60 °C for 3 s. Melting curve analysis confirmed that the PCR product did not contain non-specific by-products. TATA-binding protein (Tbp) was used as the internalnormalizing control for the mRNA. The cycle