Isolering av livskraftig flercelliga körtlar från Tissue av köttätande växt, Nepenthes

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Rottloff, S., Mithöfer, A., Müller, U., Kilper, R. Isolation of Viable Multicellular Glands from Tissue of the Carnivorous Plant, Nepenthes. J. Vis. Exp. (82), e50993, doi:10.3791/50993 (2013).

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Abstract

Materials

Name Company Catalog Number Comments
RNAqueous-Micro Kit Life Technologies GmbH, Germany AM1931 Isolation of total RNA, 50 samples
Dynabeads mRNA DIRECT Micro Kit Life Technologies GmbH, Germany 61011 5 ml Dynabeads Oligo (dT)25
DynaMag-2 magnet Life Technologies GmbH, Germany 12321D
SuperScript CellsDirect cDNA Synthesis System Life Technologies GmbH, Germany 18080-200 25 reactions
RNaseOUT Life Technologies GmbH, Germany 10777-019 Rnase inhibitor, 5,000 units
Random Hexamers (0.4 μg/μl) QIAGEN GmbH, Germany 79236 100 μl
10 mM dNTP Mix, molecular biology grade Fermentas GmbH, Germany R0192 1 ml
Taq DNA polymerase, (recombinant) Fermentas GmbH, Germany EP0402 Supplied with 10x Taq buffer (with KCl or (NH4)2SO4) and 25 mM MgCl2
TURBO DNase Life Technologies GmbH, Germany AM2238 1,000 units
Agarose Carl Roth GmbH Co. Germany T846.3
GeneRuler Low Range DNA ladder, ready-to-use Fermentas GmbH, Germany SM1193 50 μg
Brilliant II SYBR Green QPCR Master Mix Agilent Technologies, Inc., USA 600828 Single kit
RNaseZAP Sigma-Aldrich, Taufkirchen, Germany R2020
Razor blade Carl Roth GmbH Co., Germany CK08.1
Tweezers Carl Roth GmbH Co., Germany 2801.1 / 2855.1
GE NanoVue spectrophotometer GE Healthcare Europe GmbH, Germany 28-9569-62
Mastercycler gradient Eppendorf AG, Hamburg, Germany 5331 000.010
Mx3000P Real-Time PCR System Stratagene, USA 401403
Agagel Standard Horizontal Gel Electrophoresis Chamber Biometra Discontinued
Micromanipulator aura optik gmbh aureka Equipped with a microforceps
Stereomicroscope Carl Zeiss Microscopy, Jena, Germany SteREO Lumar.V12 Equipped with an AxioCam
Lumar Filter set 01 Carl Zeiss, Microscopy, Jena, Germany 485001-0000-000 UV filter
Lumar Filter set 09 Carl Zeiss, Microscopy, Jena, Germany 485009-0000-000 non-UV filter

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References

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  17. Mithöfer, A., Boland, W. Plant defense against herbivores: Chemical aspects. Ann. Rev. Plant Biol. 63, 431-450 (2012).
  18. Rottloff, S., Stieber, R., Maischak, H., Turini, F. G., Heubl, G., Mithöfer, A. Functional characterization of a class III acid endochitinase from the traps of the carnivorous pitcher plant genus, Nepenthes. J. Exp. Bot. 62, 4649-4647 (2011).

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