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- Hilson, P., et al.
(author)
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Versatile gene-specific sequence tags for Arabidopsis functional genomics : Trancript profiling and reverse genetics applications
- 2004
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In: Genome Research. - : Cold Spring Harbor Laboratory. - 1088-9051 .- 1549-5469. ; 14:10B, s. 2176-2189
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Journal article (peer-reviewed)abstract
- Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics.
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- Yafa, C., et al.
(author)
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Development of an ombrotrophic peat bog (low ash) reference material for the determination of elemental concentrations
- 2004
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In: Journal of Environmental Monitoring. - : Royal Society of Chemistry (RSC). - 1464-0325 .- 1464-0333. ; 6:493, s. 501-
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Journal article (peer-reviewed)abstract
- Given the increasing interest in using peat bogs as archives of atmospheric metal deposition, the lack of validated sample preparation methods and suitable certified reference materials has hindered not only the quality assurance of the generated analytical data but also the interpretation and comparison of peat core metal profiles from different laboratories in the international community. Reference materials play an important role in the evaluation of the accuracy of analytical results and are essential parts of good laboratory practice. An ombrotrophic peat bog reference material has been developed by 14 laboratories from nine countries in an inter-laboratory comparison between February and October 2002. The material has been characterised for both acid-extractable and total concentrations of a range of elements, including Al, As, Ca, Cd, Cr, Cu, Fe, Hg, Mg, Mn, Na, Ni, P, Pb, Ti, V and Zn. The steps involved in the production of the reference material (i.e. collection and preparation, homogeneity and stability studies, and certification) are described in detail.
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- Mironov, D, et al.
(author)
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Radiatively driven convection in ice-covered lakes: Observations, scaling, and a mixed layer model
- 2002
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In: Journal of Geophysical Research. - 2156-2202. ; 107:C4
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Journal article (peer-reviewed)abstract
- Penetrative convection is discussed where the instability is driven by radiative heating of water below the temperature of maximum density. Convection of this type occurs in ice-covered freshwater lakes in late spring, when the snow cover vanishes and solar radiation is absorbed beneath the ice cover. The vertical temperature structure, bulk mixed layer scaling, and mixed layer deepening are examined for a number of temperate and polar lakes. A bulk mixed layer scaling for this type of convection is based on energy arguments underlying the classical Deardorff convective scaling. The depth of the convective layer serves as an appropriate length scale. However, a modified scale that takes account of the energetics of a distributed radiation source term replaces the surface buoyancy flux velocity scale used by Deardorff. The scaling compares favorably with large-eddy simulations of turbulence kinetic energy (TKE) and with both observations and large-eddy simulations of the TKE dissipation rate. Mixed layer deepening is simulated with a model of convection beneath lake ice. The model describes the structure of the stably stratified layer just beneath the ice with a stationary solution to the heat transfer equation; the structure of the entrainment layer is parameterized with a zero-order jump approach. The entrainment equation is derived from the mixed layer TKE budget and bulk mixed layer scaling. Entrainment regimes characteristic of convection beneath ice are analyzed. It is shown that if the Deardorff convective velocity scale is replaced with a scale incorporating the distributed buoyancy flux, the entrainment equation describing atmospheric and oceanic convective boundary layers also applies beneath the ice. Model predictions compare well with data from observations in a number of lakes. We propose and compare with observations an extension of the mixed layer model that allows for the inclusion of salinity. Although the salt concentration is low in most temperate and polar lakes, its dynamical effect can be significant close to the temperature of maximum density.
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