Download Stable Isotope Probing and Related Technologies by J. Colin Murrell PDF

By J. Colin Murrell

The perfect start line for investigating, constructing, and enforcing strong isotope technologies.

  • Guides researchers via easy, verified, and confirmed protocols together with DNA, RNA, protein, and phospholipid fatty acid (PLFA) SIP, from suggestion and background via exact technique, troubleshooting, and interpretation to optimum and destiny uses.
  • Explores vital and rising functions of SIP in environmental microbiology, starting from bioremediation and gene mining to carbon monitoring and intestine microflora functionality.
  • Examines explorations of extra based isotope labeling applied sciences reminiscent of Raman-FISH, NanoSIMS, and isotope arrays.
  • Serves as a priceless source for environmental microbiology scholars and researchers and genomics, biotechnology, and scientific microbiology execs.

Show description

By J. Colin Murrell

The perfect start line for investigating, constructing, and enforcing strong isotope technologies.

  • Guides researchers via easy, verified, and confirmed protocols together with DNA, RNA, protein, and phospholipid fatty acid (PLFA) SIP, from suggestion and background via exact technique, troubleshooting, and interpretation to optimum and destiny uses.
  • Explores vital and rising functions of SIP in environmental microbiology, starting from bioremediation and gene mining to carbon monitoring and intestine microflora functionality.
  • Examines explorations of extra based isotope labeling applied sciences reminiscent of Raman-FISH, NanoSIMS, and isotope arrays.
  • Serves as a priceless source for environmental microbiology scholars and researchers and genomics, biotechnology, and scientific microbiology execs.

Show description

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Extra info for Stable Isotope Probing and Related Technologies

Example text

L. Blackall. 2005. Investigation of an acetate-fed denitrifying microbial community by stable isotope probing, full-cycle rRNA analysis, and fluorescent in situ hybridizationmicroautoradiography. Appl. Environ. Microbiol. 71: 8683–8691. Gorham, E. 1991. Northern peatlands: role in the carbon cycle and probable responses to climatic warming. Ecol. Appl. 1:182–195. Handelsman, J. 2005. Metagenomics: application of genomics to uncultured microorganisms. Microbiol. Mol. Bio. Rev. 68:669–685. , M. R.

While the requirements with respect to quantity of total RNA are relatively modest when compared to magnetic bead probe capture (1 mg compared with 10 mg), there are some humic-­r ich environments with low cell numbers and activity from which it is not possible to retrieve RNA of the quantity and quality required for SIP analysis (MacGregor and Amann, 2006). Additionally, because RNA is more prone to degradation through high temperature, high pH, or nucleases than DNA is, precautions (low temperature and pH) must be taken to maintain the integrity of RNA from the point of extraction through to the production of cDNA by reverse transcription.

Despite the fact that no evidence was obtained indicating the mRNA templates increased in density over time or were more abundant in high-­density gradient fractions, it is clear that RNA-­SIP also provides access to the labeled metatranscriptome. PROBLEMS AND PITFALLS OF THE TECHNIQUE; ADVANTAGES AND DISADVANTAGES RNA-­SIP has a number of advantages and disadvantages in comparison with alternative methods designed to achieve a link between microbial community function and phylogeny. RNA-­SIP, like DNA-­SIP, represents a method that addresses the open question, Which taxonomic group is responsible for consumption of substrate Y?

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