Authors: K. Drese, F. von Germar, T. Roesser, T. Hansen-Hagge, J. Bullema and P. Bolt
Affilation: TNO Science and Industry, Netherlands
Pages: 235 - 238
Keywords: cells, lysis, DNA
Many diagnostic or analytical tasks are based on detection of specific amplificates of DNA or RNA. Lab-on-Chip systems are developed for such tests because often only small sample volumes and analyte concentrations are available. A true Lab-on-Chip system should start the process at the earliest possible stage, ideally using whole blood, smears or tissue samples. To get access to DNA/RNA, the cells have to be disintegrated. A study was carried out into the efficacy of different lysis methods, when implemented on a Lab-on-Chip, with regards to the yield of amplificable DNA. Investigated methods included chemical lysis with guanidinium SCN, biochemical lysis with proteinase K, thermal lysis, use of ultrasound as well as electrical fields, a strong pH gradient inducted by localised electrolysis and mechanical lysis. Test were carried out with a standard chip design, which could be adapted to the specific method. Purity as well as concentration of released genomic DNA was inspected by agarose gel electrophoresis. Additionally, the DNA was amplified by qPCR to test the amount of amplificable DNA. Moreover, loss of DNA during lysis and purification steps was monitored by quantification of bacteriophage lambda DNA added to the sample before lysis.