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  • Cold Spring Harbor Laboratory  (6)
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  • Cold Spring Harbor Laboratory  (6)
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  • 1
    Online-Ressource
    Online-Ressource
    Cold Spring Harbor Laboratory ; 2012
    In:  Cold Spring Harbor Protocols Vol. 2012, No. 3 ( 2012-03), p. pdb.prot067975-
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2012, No. 3 ( 2012-03), p. pdb.prot067975-
    Kurzfassung: This article describes an immunostaining protocol for Ectocarpus that was optimized for the detection of tubulin but could be used with any suitable antibody. Ectocarpus has small but relatively transparent cells and the uniseriate filaments can be grown directly attached to the surface of microscope slides. These features make Ectocarpus particularly suitable for high resolution imaging approaches, both in vivo or after fixation. All incubations described below are carried out on a platform shaker at room temperature. Use high-quality microscope slides to avoid imperfections in the glass that can be a problem for confocal laserscan microscopy analysis.
    Materialart: Online-Ressource
    ISSN: 1940-3402 , 1559-6095
    Sprache: Englisch
    Verlag: Cold Spring Harbor Laboratory
    Publikationsdatum: 2012
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 2
    Online-Ressource
    Online-Ressource
    Cold Spring Harbor Laboratory ; 2012
    In:  Cold Spring Harbor Protocols Vol. 2012, No. 3 ( 2012-03), p. pdb.prot067959-
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2012, No. 3 ( 2012-03), p. pdb.prot067959-
    Kurzfassung: This article describes how to obtain isolated cells with no surrounding cell wall by enzymatic digestion of Ectocarpus filaments. The resultant protoplasts are totipotent and regenerate to produce individual algae under appropriate culture conditions. The yield of protoplasts and their capacity to regenerate are highly dependent on the Ectocarpus strain used, the stage of the life cycle, and the culture conditions. The highest yields are obtained with young gametophyte filaments cultivated at low density. The naked, wall-less cells produced by this protocol can be used for several applications, including studies of cell wall regeneration, investigation of the role of the cell wall in determining cell fate, and as a source of naked cells for the development of methods for introducing diverse molecules into the cell.
    Materialart: Online-Ressource
    ISSN: 1940-3402 , 1559-6095
    Sprache: Englisch
    Verlag: Cold Spring Harbor Laboratory
    Publikationsdatum: 2012
    Standort Signatur Einschränkungen Verfügbarkeit
    BibTip Andere fanden auch interessant ...
  • 3
    Online-Ressource
    Online-Ressource
    Cold Spring Harbor Laboratory ; 2012
    In:  Cold Spring Harbor Protocols Vol. 2012, No. 2 ( 2012-02), p. pdb.emo065821-
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2012, No. 2 ( 2012-02), p. pdb.emo065821-
    Kurzfassung: The brown algae are an interesting group of organisms from several points of view. They are the dominant organisms in many coastal ecosystems, where they often form large, underwater forests. They also have an unusual evolutionary history, being members of the stramenopiles, which are very distantly related to well-studied animal and green plant models. As a consequence of this history, brown algae have evolved many novel features, for example in terms of their cell biology and metabolic pathways. They are also one of only a small number of eukaryotic groups to have independently evolved complex multicellularity. Despite these interesting features, the brown algae have remained a relatively poorly studied group. This situation has started to change over the last few years, however, with the emergence of the filamentous brown alga Ectocarpus as a model system that is amenable to the genomic and genetic approaches that have proved to be so powerful in more classical model organisms such as Drosophila and Arabidopsis .
    Materialart: Online-Ressource
    ISSN: 1940-3402 , 1559-6095
    Sprache: Englisch
    Verlag: Cold Spring Harbor Laboratory
    Publikationsdatum: 2012
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 4
    Online-Ressource
    Online-Ressource
    Cold Spring Harbor Laboratory ; 2012
    In:  Cold Spring Harbor Protocols Vol. 2012, No. 3 ( 2012-03), p. pdb.prot067967-
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2012, No. 3 ( 2012-03), p. pdb.prot067967-
    Kurzfassung: For some applications, such as genome sequencing and high-throughput genotyping with multiple markers, it is necessary to use high-quality genomic DNA. This article describes how to obtain several micrograms of high-quality, cesium chloride-purified DNA from 1 g of Ectocarpus filaments. We also recommend using DNA of this quality for quantitative RT–PCR control reactions. However, simpler, more rapid, kit-based methods are preferable for experiments that involve the treatment of large numbers of individuals, such as genotyping large populations with a small number of markers or PCR screening of large populations.
    Materialart: Online-Ressource
    ISSN: 1940-3402 , 1559-6095
    Sprache: Englisch
    Verlag: Cold Spring Harbor Laboratory
    Publikationsdatum: 2012
    Standort Signatur Einschränkungen Verfügbarkeit
    BibTip Andere fanden auch interessant ...
  • 5
    Online-Ressource
    Online-Ressource
    Cold Spring Harbor Laboratory ; 2012
    In:  Cold Spring Harbor Protocols Vol. 2012, No. 2 ( 2012-02), p. pdb.prot067934-
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2012, No. 2 ( 2012-02), p. pdb.prot067934-
    Kurzfassung: This article describes the standard procedure for growing Ectocarpus in the laboratory. The culture is started with partheno-sporophyte (or sporophyte) filaments because this is the stage that is usually maintained in strain collections. The standard medium is Provasoli-enriched natural seawater (PES), but Ectocarpus can also be grown in artificial seawater, which allows more precise control over the culture conditions. The algae can be cultivated either in plastic Petri dishes or in 10-L bottles with bubbling, if large amounts of biomass are required. Standard growth conditions are 13°C with a 12h/12h d/night cycle and 20 µmol photons m −2 s −1 irradiance using daylight-type fluorescent tubes. All manipulations of Ectocarpus cultures should be performed in a clean environment (if possible, under a laminar flow hood). Forceps should be dipped in ethanol and allowed to dry under the hood.
    Materialart: Online-Ressource
    ISSN: 1940-3402 , 1559-6095
    Sprache: Englisch
    Verlag: Cold Spring Harbor Laboratory
    Publikationsdatum: 2012
    Standort Signatur Einschränkungen Verfügbarkeit
    BibTip Andere fanden auch interessant ...
  • 6
    Online-Ressource
    Online-Ressource
    Cold Spring Harbor Laboratory ; 2012
    In:  Cold Spring Harbor Protocols Vol. 2012, No. 2 ( 2012-02), p. pdb.prot067942-
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2012, No. 2 ( 2012-02), p. pdb.prot067942-
    Kurzfassung: This article describes a procedure for conducting crosses between different strains of Ectocarpus . Crossing gametophytes to obtain the sporophyte generation is the most technically challenging stage of this process because diploid sporophytes have to be distinguished from the haploid partheno-sporophytes that result from the parthenogenetic germination of unfused gametes. This requires careful monitoring of the progeny of the genetic cross until they have developed sufficiently to be transferred to a separate Petri dish. Genetic crosses allow several classical genetic methodologies to be applied in Ectocarpus , including allelic complementation tests, backcrosses, combination of different genetic mutations, and outcrosses to create mapping populations.
    Materialart: Online-Ressource
    ISSN: 1940-3402 , 1559-6095
    Sprache: Englisch
    Verlag: Cold Spring Harbor Laboratory
    Publikationsdatum: 2012
    Standort Signatur Einschränkungen Verfügbarkeit
    BibTip Andere fanden auch interessant ...
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