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Biochemistry

RNA Chromatin Immunoprecipitation (RNA-ChIP) in Caenorhabditis elegans Authors:  Germano Cecere and Alla Grishok, date: 12/20/2014, view: 6334, Q&A: 0
The RNA chromatin immunoprecipitation assay (RNA-ChIP) allows detection and quantification of RNA–protein interactions using in vivo cross-linking with formaldehyde followed by immunoprecipitation of the RNA–protein complexes. Here we describe the RNA–ChIP protocol that we have adapted for Caenorhabditis elegans (C. elegans ...
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Microbiology

Hepatitis C virus Cell-to-cell Spread Assay Authors:  Naina Barretto and Susan L. Uprichard, date: 12/20/2014, view: 4740, Q&A: 0
Hepatitis C virus (HCV) can infect naïve cells via entry of “cell-free” extracellular virus or direct “cell-to-cell” transmission. Here, we describe an assay for detecting HCV cell-to-cell transmission, using a non-growing cell culture system that avoids confounding effects of cell growth. The assay consists of infecting a small number of cells in ...
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Fluorescence-linked Antigen Quantification (FLAQ) Assay for Fast Quantification of HIV-1 p24Gag Authors:  Marianne Gesner, Mekhala Maiti, Robert Grant and Marielle Cavrois, date: 12/20/2014, view: 4479, Q&A: 1
The fluorescence-linked antigen quantification (FLAQ) assay allows a fast quantification of HIV-1 p24Gag antigen. Viral supernatant are lysed and incubated with polystyrene microspheres coated with polyclonal antibodies against HIV-1 p24Gag and detector antibodies conjugated to fluorochromes (Figure 1). After washes, the ...
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Plant Science

Cyclohexane Diamine Tetraacetic Acid (CDTA) Extraction of Plant Cell Wall Pectin Authors:  Gerit Bethke and Jane Glazebrook, date: 12/20/2014, view: 5316, Q&A: 0
The goal of this procedure is to extract pectin from plant cell walls. Pectins are galacturonic acid containing polymeric sugars that are important components of plant cell walls. Various procedures aimed at studying plant cell wall components require the extraction of pectin. Pectin is synthesized in the Golgi apparatus in a highly esterified ...
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Virus-induced Gene Silencing (VIGS) in Phalaenopsis Orchids Authors:  Ming-Hsien Hsieh, Zhao-Jun Pan, Hsin-Hung Yeh and Hong-Hwa Chen, date: 12/20/2014, view: 5751, Q&A: 0
This is a protocol to produce stable silencing efficacy and efficiency for VIGS using CymMV as a silencing vector for floral functional genomics in Phalaenopsis orchids. This protocol is established based on a method created by Lu et al. (2007), and then modified by Hsieh et al. (2013a; 2013b), Lu et al. (2012) ...
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Imaging and Measurement of Nanomechanical Properties within Primary Xylem Cell Walls of Broadleaves Authors:  Jaroslav Ďurkovič, Monika Kardošová and Rastislav Lagaňa, date: 12/20/2014, view: 5549, Q&A: 0
A technique of atomic force microscopy (AFM) called PeakForce quantitative nanomechanical mapping (PeakForce QNM) is an efficient tool for the quantitative mechanobiological imaging of fibrillar aggregate, human epidermal cell and woody plant cell wall topography (Sweers et al., 2011; Heu et al., 2012; Ďurkovič et al., ...
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Trichome Isolation and Integrity Test from Brassica villosa and Other Species Authors:  Naghabushana Nayidu, Peta Bonham-Smith and Margaret Y. Gruber, date: 12/20/2014, view: 4577, Q&A: 0
The outward growths from one or more epidermal cells are well known as trichomes (plant hair cells) (Levin, 1973; Mathur, 2006). Preparation of pure intact non-glandular trichomes from trichome-rich Brassica villosa depended on rendering the trichomes sufficiently stiff to dislodge them from the leaves in an undamaged state, which can be ...
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Agrobacterium-mediated Transformation of Mature Ginseng Embryos Authors:  Ok Ran Lee, Jin Haeng Han and Yoon-Ri Kim, date: 12/20/2014, view: 5136, Q&A: 0
Ginseng refers to species within the genus Panax and is a slow-growing perennial herb from the Araliaceae family. The most widely used Panax species is Panax ginseng Meyer (Korean ginseng). Panax japonicus (Japanese ginseng), Panax notoginseng (Chinese ginseng), and Panax quinquefolium (American ...
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Localization and Topology of Thylakoid Membrane Proteins in Land Plants Authors:  Salar Torabi, Magdalena Plöchinger and Jörg Meurer, date: 12/20/2014, view: 5514, Q&A: 0
Thylakoids are a formation of flattened membrane vesicles and protein complexes found in cyanobacteria, algae and plants. In the chloroplasts of land plants the thylakoid membrane systems form a network of densely packed stacks called grana lamellae, which are connected by unstacked stroma lamellae. Photosystem II is mainly localized in the ...
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Polysomal-mRNA Extraction from Arabidopsis by Sucrose-gradient Separation Authors:  Shih-Yun Lin and Guang-Yuh Jauh, date: 12/20/2014, view: 5256, Q&A: 0
mRNAs surrounded by polysomes are ready for translation into proteins (Warner et al., 1963); these mRNAs are defined as polysomal-mRNAs (Mustroph et al., 2009). The process is affected by various growth conditions or surrounding situations. Microarray analysis is a powerful tool for detecting genome-wide gene expression. ...
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