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  Air-sampled Filter Analysis for Endotoxins and DNA Content

Lang-Yona, N., Mazar, Y., Pardo, M., & Rudich, Y. (2016). Air-sampled Filter Analysis for Endotoxins and DNA Content. Journal of visualized experiments, (109): UNSP e53444. doi:10.3791/53444.

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 Creators:
Lang-Yona, N.1, Author           
Mazar, Yinon2, Author
Pardo, Michal2, Author
Rudich, Yinon2, Author
Affiliations:
1Multiphase Chemistry, Max Planck Institute for Chemistry, Max Planck Society, ou_1826290              
2external, ou_persistent22              

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 Abstract: Outdoor aerosol research commonly uses particulate matter sampled on filters. This procedure enables various characterizations of the collected particles to be performed in parallel. The purpose of the method presented here is to obtain a highly accurate and reliable analysis of the endotoxin and DNA content of bio-aerosols extracted from filters. The extraction of high molecular weight organic molecules, such as lipopolysaccharides, from sampled filters involves shaking the sample in a pyrogen-free water-based medium. The subsequent analysis is based on an enzymatic reaction that can be detected using a turbidimetric measurement. As a result of the high organic content on the sampled filters, the extraction of DNA from the samples is performed using a commercial DNA extraction kit that was originally designed for soils and modified to improve the DNA yield. The detection and quantification of specific microbial species using quantitative polymerase chain reaction (q-PCR) analysis are described and compared with other available methods.

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 Dates: 2016
 Publication Status: Published online
 Pages: -
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 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000374636300013
DOI: 10.3791/53444
 Degree: -

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Title: Journal of visualized experiments
  Other : Journal of visualized experiments: JoVE
  Abbreviation : J. Vis. Exp.
Source Genre: Journal
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Publ. Info: Rockville Pike, Bethesda MD : JoVE
Pages: - Volume / Issue: (109) Sequence Number: UNSP e53444 Start / End Page: - Identifier: ISSN: 1940-087X
CoNE: https://pure.mpg.de/cone/journals/resource/1940087X