Search
  • Volume 7, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Cell Biology

Micrococcal Nuclease (MNase) Assay of Arabidopsis thaliana Nuclei Authors:  Laia Armengot and Jordi Moreno-Romero, date: 04/05/2013, view: 8501, Q&A: 2
Micrococcal nuclease (MNase) is able to produce double-strand breaks within nucleosome linker regions. The efficiency of MNase digestion depends on the degree of chromatin compaction, being more easily digested the regions of less compacted chromatin. The MNase protocol described here can be used to asses changes in the chromatin structure of ...
More >>

Immunology

Allergen Sensitization and Challenge to Ovalbumin Authors:  François Daubeuf, Laurent Reber and Nelly Frossard, date: 04/05/2013, view: 8129, Q&A: 0
This protocol describes the sensitization and challenge of mice with ovalbumin for use as an acute murine model of asthma. This protocol induces reproducible airway inflammation and remodelling, and bronchial hyperresponsiveness to methacholine as measured by barometric plethysmography, as well as by the Flexivent® technique in ...
More >>
Measurement of Airway Responsiveness in the Anesthetized Mouse Authors:  François Daubeuf, Laurent Reber and Nelly Frossard, date: 04/05/2013, view: 6636, Q&A: 0
Airway hyperresponsiveness to methacholine is an important characteristic of asthma. Many devices can be used to measure airway responsiveness in the mouse but it is well established that the invasiveness of a measurement technique is correlated with its precision and reproducibility. This protocol describes how to measure airway responses to ...
More >>

Microbiology

Neutral Red Assay for Murine Norovirus Replication and Detection in a Mouse Authors:  Mariam Bernadette González-Hernández, Jeffrey William Perry and Christiane E. Wobus, date: 04/05/2013, view: 7249, Q&A: 0
Neutral red (NR) is a dye that must be actively imported into the cell, and, therefore, the dye has been used for decades to selectively stain living cells. In addition, NR can also be incorporated into virus particles, although the mechanism behind this is poorly understood. Once encapsulated into the virion, NR, a light sensitive dye, can be ...
More >>
HIV-1 Single Cycle Infection Authors:  Ferdinand Roesch and Olivier Schwartz, date: 04/05/2013, view: 5371, Q&A: 0
The role of a viral or cellular protein on HIV-1 infection can sometimes be difficult to assess in a system where the virus is able to replicate for several cycles. Indeed, some effects are only observable at early time points, and can be masked (or on the contrary, artificially increased) after several rounds of viral replication. Therefore, to ...
More >>
Quantification of HIV-1 DNA Authors:  Ferdinand Roesch and Olivier Schwartz, date: 04/05/2013, view: 5741, Q&A: 0
The reverse transcription (RT) reaction is a critical step in HIV-1 life cycle. It is very strongly regulated and the target of several restriction factors (TRIM5α, APOBECs, SAMHD1, etc.). The progress of reverse transcription can be followed by measuring viral DNA by quantitative PCR (qPCR). This method is sensitive enough to allow detection of ...
More >>
Fusarium Virulence Assay on Wheat and Barley Seedlings Authors:  Lorenzo Covarelli, Donald Gardiner, Giovanni Beccari and Paul Nicholson, date: 04/05/2013, view: 6445, Q&A: 0
Fusarium root and crown rot is a very important complex disease of small grain cereals worldwide which may lead to very high yield losses. Traditional virulence assays are time consuming and often require plants to be grown in greenhouses or climatic chambers in soil. We describe a rapid laboratory assay for assessing such a disease in wheat and ...
More >>

Molecular Biology

Western Analysis of Histone Modifications (Aspergillus nidulans) Authors:  Alexandra Soukup and Nancy P. Keller, date: 04/05/2013, view: 6129, Q&A: 0
Western blotting allows for the specific detection of proteins and/or modifications of proteins by an antibody of interest. This protocol utilizes a crude nuclei extraction protocol for Aspergillus nidulans to enrich for histones and other nuclear proteins prior to gel electrophoresis. Post translational modifications of histones may ...
More >>
Northern Blot of tRNA in Yeast Author:  Yuehua Wei, date: 04/05/2013, view: 7015, Q&A: 0
tRNAs are small RNAs around 70-90 nt. tRNAs are different from many other small RNAs in that they are very abundant, which makes it difficult to study their transcriptional regulation by traditional northern blot. Traditional Northern blot involves incorporation of radioactive nucleotides through polymerization, however, tRNA is too short for ...
More >>
Metabolic Labeling of Yeast RNA with Radioactive Uracil Author:  Yuehua Wei, date: 04/05/2013, view: 5136, Q&A: 0
To examine gene expression, Northern blot or Real-Time PCR can be used to detect low abundant RNA such as mRNA. However, for high abundant RNAs such as rRNA and tRNA, Northern blot will not be able to discriminate the newly synthesized RNA from total RNA. Therefore, metabolic labeling is necessary to evaluate the expression of rRNA and tRNA genes. ...
More >>

Neuroscience

Fear Conditioning Assay in Mouse Authors:  Melissa Wang and Isabel A. Muzzio, date: 04/05/2013, view: 6151, Q&A: 0
The study of fear memory is important for understanding various anxiety disorders in which patients experience persistent recollections of traumatic events. These memories often involve associations of contextual cues with aversive events; consequently, Pavlovian classical conditioning is commonly used to study contextual fear learning. A form of ...
More >>

Plant Science

Northwestern Blot of Protein-RNA Interaction from Young Rice Panicles Authors:  Saminathan Thangasamy and Guang-Yuh Jauh, date: 04/05/2013, view: 7133, Q&A: 0
The northwestern assay is employed to study the interaction between protein and RNA. The RNA binding proteins tend to bind to different kinds of RNA through either known domains or unknown sequences of proteins. Rice LGD1 recombinant protein, a grass-specific novel protein with RNA binding sequences in its C-terminal, was used to probe its ...
More >>