DNA quantification Sample Clauses

DNA quantification. The extracted DNA concentration was measured using the Qubit® 2.0 Fluorometer DS DNA BR ASSAY KIT provided by Invitrogen (Life Technology, Paisley, UK). Two Qubit Assay tubes of 0.5 μl for the standards (S1 & S2) and one for each sample were prepared. All tubes were clearly labelled. A master working solution was prepared by mixing 199 μl/sample of the Qubit Buffer with 1 μl/sample of the Qubit Reagent. 200 μl of the working solution were prepared for each standard and sample. In each numbered sample tube 199 μl of the working solution were mixed with 1 μl of DNA to a final volume of 200 μl. Tubes S1 and S2 were prepared by mixing 190 μl of the working solution with 10 μl of the respective standard to obtain a final volume of 200 μl. All samples were mixed by vortexing for 2 minutes and then incubated for 2 minutes at room temperature. Standard samples were kept in a non- transparent box for the same incubation time. Standards S1 and S2 were first inserted into the fluorometric device followed by each sample individually to record DNA concentration in ng/μl and the 260/280 ratio for potential sample contamination.

DNA quantification. High quality DNA concentration (20 ng/μl, A260/A280 1.8-2.0) was confirmed with the Qubit