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Department of Microbiology and Immunology, University of British Columbia - UBC
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Walsh, D. A., Zaikova, E., Hallam, S. J. Small Volume (1-3L) Filtration of Coastal Seawater Samples. J. Vis. Exp. (28), e1163, doi:10.3791/1163 (2009).
代表性的成果:每1升的水样产生一个Sterivix过滤器,其中包含生物质能大于0.22微米。这是存储在随后的DNA提取和分析裂解缓冲液。最终结果由一个Sterivix过滤裂解缓冲液中,每存储1升的瓶子,并准备提取DNA。该过滤器包含生物质能大于0.22微米。过滤器可以储存在-80 ° C之前的eDNA提取。
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当务之急是盐水中删除任何挥之不去的痕迹,否则将腐蚀设备后,使用湿毛巾擦拭长椅和泵。虽然过程很简单,时间要求,可广泛取决于生物质的浓度或在被筛选的样本碎片。
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我们要感谢支持沿海和公海水域低氧气地区正在进行的研究创新,不列颠哥伦比亚省知识发展基金和国家科学和工程研究理事会(NSERC),加拿大的加拿大基金会。提高妇女地位司的支持由NSERC,Killam和图拉基金会资助的微生物多样性和进化中心奖学金。
| Name | Company | Catalog Number | Comments |
| Peristaltic pump | Masterflex (Cole Palmer) | 77913-20, 77200-62, 77200-04 | |
| Tygon LFL tubing | Masterflex (Cole Palmer) | 06429-24 | For filtration apparatus |
| Hose clamps | Cole-Parmer | 0683204 | For filtration apparatus |
| 1/4” x 3/16” pipe adaptors | Cole-Parmer | 31702-20 | For filtration apparatus |
| Polypropylene prefilter housing | ADVANTEC | 501200 | For filtration apparatus |
| Metal luer fitting (Hose end to Male luer lock) | Cole-Parmer | 31507-27 | For filtration apparatus |
| 2“ Magnets | For filtration apparatus | ||
| 3 port carboy caps | For filtration apparatus | ||
| GV 0.22 μm STERIVIX filters | EMD Millipore | SVGV01R5 | |
| 47 mm diameter 2.7 μm GF/D filters | Whatman, GE Healthcare | 1823047 | |
| Lysis buffer | 0.75 M sucrose, 40 mM EDTA, 50 mM Tris (pH 8.3) | ||
| 0.1 % HCl | For post-filtration rinse | ||
| Autoclaved water | For post-filtration rinse |
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ReplyPosted by: StephenJuly 11, 2011, 10:23 PM