The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

Recommend to Librarian

Automatic Translation

This translation into Chinese was automatically generated through Google Translate.
English Version | Other Languages

 JoVE General

沿海海水样品的过滤体积小(1 - 3L)

, ,

Department of Microbiology and Immunology, University of British Columbia - UBC

You must be subscribed to JoVE to access this content.

This article is a part of   JoVE General. If you think this article would be useful for your research, please recommend JoVE to your institution's librarian.

Recommend JoVE to Your Librarian

Current Access Through Your IP Address

You do not have access to any JoVE content through your current IP address.

IP: 107.20.7.65, User IP: 107.20.7.65, User IP Hex: 1796474689

Current Access Through Your Registered Email Address

You aren't signed into JoVE. If your institution subscribes to JoVE, please or create an account with your institutional email address to access this content.

 

Video Article Chapters

Cite this Article: 沿海海水样品的过滤体积小(1 - 3L)

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).

Abstract: 沿海海水样品的过滤体积小(1 - 3L)

工作流程开始与下游微生物群落的沿海海域,养分和微量气体分析的集合。对于今天的示范样本收集从甲板HMS施德Saanich入口经营。此视频文件体积小(约1升)过滤的微生物生物量的水柱。该协议是延长大体积取样协议如前所述,一个主要的区别:在这里,有没有预过滤步骤,让所有大小类生物质收集到0.22微米的过滤器切断。样品收集这种方式,是理想的核酸分析。设置,过滤和清理的每个步骤需时约20-30分钟。如果同时使用两个蠕动泵,最多8个样品,同时可过滤。为了防止生物膜之间的采样人次的形成,所有的过滤设备,必须用稀盐酸和去离子水冲洗,高压灭菌后立即使用。

Protocol: 沿海海水样品的过滤体积小(1 - 3L)

  1. 设置过滤装置
    1. 过滤前,先擦去从满瓶凝露和权衡
    2. 将泵一侧的高压灭菌器斌,其他废物瓶。
    3. 然后将一个完整的样瓶,在蒸压斌;但是,不要删除瓶盖。
    4. 通过蠕动泵管,使油管露儿烧瓶中。
    5. 使用镊子,取出25毫升无菌吸管棉花,和吸管连接油管的另一端。
  2. 海水过滤
    1. 要开始过滤海水,打开瓶盖,插入吸管。打开泵和流动速度设置为1。观察,以确保水是被抽在正确的方向。
    2. 允许〜100毫升的水,穿过整个油管。
    3. 关闭泵和一个Sterivex过滤器附着在油管的自由端男性露儿装修。
    4. 继续筛选,直到没有水在瓶左。
  3. 储存过滤器
    1. 要存储的过滤器,首先驱逐任何剩余的水30-60毫升注射器。
    2. 用封口膜密封的Sterivex过滤器的底部。使用吸液管,添加1.8毫升裂解液过滤器,保持在后来试剂的过滤器〜200 UL空间。
    3. 然后,用小块的封口膜的Sterivex过滤器的顶部密封,标签的日期和样品标识过滤器。
    4. 商店标记的前50毫升猎鹰管过滤器在-80 ° C
  4. 清理
    1. 25毫升吸管插入含有0.1%的盐酸瓶,泵的切换。
    2. 盐酸清洗后,用灭菌水冲洗管路。

      最后,测量和记录现在空1L瓶的质量,以计算体积的水通过过滤器。

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。

Subscription Required. Please recommend JoVE to your librarian.

Discussion: 沿海海水样品的过滤体积小(1 - 3L)

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。

Subscription Required. Please recommend JoVE to your librarian.

Disclosures: 沿海海水样品的过滤体积小(1 - 3L)

Acknowledgements: 沿海海水样品的过滤体积小(1 - 3L)

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。

Materials: 沿海海水样品的过滤体积小(1 - 3L)

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

Ask the Author: 沿海海水样品的过滤体积小(1 - 3L)

1 Comment

I'm interested about the video but unfortunately our school does not have subscription for the JoVE. Is there any other way I can access the video?

1

Reply

Posted by: StephenJuly 11, 2011, 10:23 PM

Dear Stephen, Please contact me at ward.parry@jove.com so that I can discuss your request with you. Best wishes,
Ward Parry

1.1

Reply

Posted by: Ward PArryJuly 12, 2011, 4:46 PM

Post a Question / Comment / Request

You must be signed in to post a comment. Please or create an account.

Waiting
simple hit counter