超滤在生物大分子实验中的应用.pptVIP

超滤在生物大分子实验中的应用 李明新 Millipore China LTD 2006.5 Millipore产品在生命科学领域的应用 基因组学 蛋白质组学 药物筛选 环境科学 …… Amicon - Ultra Design Amicon Ultra - Mode of Action Amicon Ultra - Mode of Action Amicon Ultra - Mode of Action Centrifugals vs. Stirred Cells Centrifugals Low cost, ease-of-use We have centrifugals to handle a full range of volumes: 1mL – 420mL Stirred cells Widely used Effective for viscous solutions Can use a 1 0.5 K membrane for peptides, etc Microcon-96 MultiScreen HTS Manifold Plasmid Kits 60 Minute processing time 10 Minute resuspension of purified BAC DNA No ethanol precipitation, no centrifugation Includes optimized sequencing protocol for BAC DNA Longer read lengths Automation compatible Less expensive Market Need Fulfilled… ‘Immobilon Western Chemiluminescent HRP Substrate’ High Sensitivity (to femtogram level) Broad detection range (nano to femtogram level) Priced less than competitors Optimized and validated with Immobilon-P Compatible with all PVDF, NC, blocking buffers Uses less antibody than low sensitivity substrates MILLIPORE 应用实例三: PCR 产物纯化 Procedure: 1. Load 400 ?l TE-buffer and PCR sample 2. Spin 15 minutes at 1000 x g 3. Add 20 ?l TE-buffer, invert spin 2 min at 1000 x g MILLIPORE MILLIPORE 注意事项 注意超滤管装置的化学兼容性 不能高温高压灭菌处理 不能超过规定的最大离心力 超滤膜一旦湿润,使用前就必须保持润湿状态 膜碎裂 MILLIPORE 储存时一般常温即可，耐受温度：4～50℃ 离心时间勿过长，否则导致膜干燥 对于浓度较高的样品，可先稀释然后再处理 抛弃型产品，一次使用效果最佳 保留好处理过的滤过液和浓缩液 合理选择超滤摸材质 最佳效果 注意事项 MILLIPORE 1 – 4 mL 4 – 30 mL 30 - 420 mL 2 – 10 L 200 mL - 2 L 100 ml - 2L Stirred Cells Centricon Plus-70 Prep/Scale Pellicon XL Amicon Ultra-15 Amicon Ultra-4 Pellicon 2 1 mL Microcon 搅拌式超滤装置 10 L MILLIPORE 搅拌式超滤装置 MILLIPORE Pressure 超滤膜 搅拌子 Concentration Polarization - Control MILLIPORE MILLIPORE For Sample volns. greater then stirred cell. The stirred cell should be filled to it’s maximum volume with remainder added to reservoir. Pressure CDS 10 Manifold MILLIPORE MILLIPOR