P0135 Efficient sample preparation of various animal and plant tissues prior to nucleic acid purification with Thermo Scientific KingFisher Duo and KingFisher Kits

Marika Suomalainen , Thermo Fisher Scientific, Vantaa, Finland
Sini Suomalainen , Thermo Fisher Scientific
Vuokko Kytoniemi , Thermo Fisher Scientific
Virpi Puro , Thermo Fisher Scientific
Arja Lamberg , Thermo Fisher Scientific
Efficient homogenization is required to release nucleic acids from cells and tissues, thus it is an essential step to ensure good DNA and RNA yield and quality. Certain sample types, especially harder tissue samples, require strong, either chemical or mechanical treatment for destroying cellular structures before nucleic acid purification process. Magnetic particle technology enables fast and effective purification of high quality nucleic acids. Thermo Scientific KingFisher Duo uses magnetic particle technology based on the use of magnetic rods with specially designed plastic consumables, BindIt software for protocol development and optimized KingFisher Kits for nucleic acid purification. KingFisher Duo enables purification of 12 samples during one run in working volumes of 20-1000 µl. Running two protocols sequentially without interruption raises throughput up to 24 samples per load. In addition, the instrument gives an option to choose working volumes up to 5 ml. We tested several different homogenization methods, such as bead beating and rotor-stator methods in addition to chemical lysis, to optimize homogenization step before the nucleic acid purification in the KingFisher Duo. We analysed performance of homogenization and purification using several different sample materials, including cells and animal or plant tissues. The purification process in the KingFisher Duo was performed using three different KingFisher Kits. The results indicate high sensitivity of the purification process. The KingFisher Duo in combination with the KingFisher Kits and new BindIt software 3.2 constitute an exceptional purification system for obtaining excellent yield and purity of nucleic acids.