动植物组织中脂肪酸检测方案(研磨机)

2010 Geno/Grinder APPLICATIONNOTE Mechanical Disruption for High-Throughput Fatty AcidExtraction from Animal Tissue Samples With kind permission of Sanofi-Aventis Germany GmbH The mechanical homogenization for extraction of fatty acids is traditionally performed with anUltra-Turrax in a buffer solution. However， both the purification and the slow sample preparationprocess in the analytical laboratory are labour-intensive. This method is no longer suitable for highsample throughput. Therefore it makes sense to combine the increased sample throughput withmechanical cell disruption. The use of 96-well micro titer plates and deep well plates is suitable forthis process. The research in biotechnology/ gene technology also often demands the handling of aconsiderable number of samples. Sample preparation is often the bottleneck in the process ofefficient analysis work. Up to now efforts to mechanically disrupt cells through grinding have beenbased on the modification of traditional ball or swing mills with an adapter for micro titer plates.The Geno/Grinder° 2000 is currently the first available mill specifically designed for cell disruption.The Geno/Grinder is a purpose-built ball mill that was originally conceived for the disruption ofplant seeds but can also be used for animal and human samples. Materials and Methods Two deep well plates (PP)， used for collecting and storing of biological materials with a1.5-2mlwells were placed next to each other in the Geno/Grinder clamp assembly. Freeze dried rat muscleor livers were placed in each plate well and spiked with 100mM KH，PO buffer at pH 2 or trichloro-acetic acid. Then two grinding balls were added. When all wells were filled， the plate was coveredwith a sealing mat. Thus prepared， the deep-well plates were clamped into the Geno/Grinder. The samples were shaken vigorously for one minute at 1700 strokes per minute. This resulted in afine， homogeneous sample dispersion within each well. Because of the vertical movement of theGeno/Grinder clamps， all wells are moved uniformly. This results in uniform and reproduciblehomogenization yields for all samples. As well as using titer plates with 96 wells it is also possible touse other formats， e.g. the"Biomek micro centrifuge 24 position tube rack" for Eppendorf Safe-Lockreaction tubes. Results and Discussions The results show that the Geno/Grinder 2000 is well suited for cell disruption of muscle and internalorgans，i.e. liver. However， the quality of the results depends on the grinding accessories. Usingseveral small Zirconium oxide beads with a diameter of 2mm does not give a satisfactory result. Thesheer force is not sufficient to grind frozen muscle tissue. In contrast to this， it was possible tosuccessfully homogenize the frozen material with stainless steel grinding balls of 4mm diameter. ：：APPLICATION NOTE SP001： Cell Disruption ：：APPARATUS：Geno/Grinder ：：APPLICATION：Cell Lysis C SPEX SamplePrep 15 Liberty St Metuchen， NJ 08840 USA Tel：732-623-0465 Fax： 732-906-2492 E-mail： Sampleprep@spexcsp.comwww.spexsampleprep.com European Headquarters SPEX CertiPrep Ltd 2 Dalston GardensStanmore， HA7 1BQ， UKTel： +44(0)208204 6656 Fax：+44(0)208204 6654 E-mail： sales@spexcertiprep.co.ukWeb： www.spexcertiprep.co.uk