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Isothermal Titration Calorimetry: A Biophysical Method to Characterize the Interaction between Label-free Biomolecules in Solution

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High-throughput YO-PRO-1 Uptake Assay for P2X7 Receptors Expressed in HEK Cells

Authors: Akira Karasawa and Toshimitsu Kawate, date: 07/20/2018, view: 178, Q&A: 0

P2X7 receptors are extracellular ATP-gated ion channels that play broad physiological and pathological roles in animals (Sluyter, 2017). Activation of P2X7 receptors lead to the opening of membrane channels permeable for small cations like Na⁺ ...

Overcoming Autofluorescence to Assess GFP Expression During Normal Physiology and Aging in Caenorhabditis elegans

Authors: Alina C. Teuscher and Collin Y. Ewald, date: 07/20/2018, view: 820, Q&A: 0

Green fluorescent protein (GFP) is widely used as a molecular tool to assess protein expression and localization. In C. elegans, the signal from weakly expressed GFP fusion proteins is masked by autofluorescence emitted from the intestinal ...

Plasmid Extract from Budding Yeast (Saccharomyces cerevisiae)

Author: Gal Haimovich, date: 07/20/2018, view: 127, Q&A: 0

Plasmids are widely used tools in yeast research. In many cases, plasmid libraries are used in genetic screens or in yeast two hybrid screens. In such cases, it is necessary to extract plasmids carrying unknown genetic elements from positive clones ...

RNA Immunoprecipitation (RIP) Sequencing of Pri-miRNAs Associated with the Dicing Complex in Arabidopsis

Authors: Ziwei Li, Songxiao Zhong, Wenye Wu and Binglian Zheng, date: 07/05/2018, view: 206, Q&A: 0

RNA immunoprecipitation (RIP) is an antibody-based technique used to map in vivo RNA-protein interactions. DBR1, an RNA debranching enzyme, is responsible for the debranching of lariat RNA, for the degradation and turnover of lariat RNAs. ...

Phenol-based Extraction of RNA from Chlamydomonas reinhardtii

Authors: Emanuel Sanz-Luque and Amaury de Montaigu, date: 07/05/2018, view: 251, Q&A: 0

RNA extraction is a basic procedure in molecular biology and a wide variety of commercial kits are available. Some of these kits have been successfully used in Chlamydomonas. However, in some cases RNA quality and quantity may be ...

In vitro Chaperone Activity Assay Using α-Amylase as Target Protein

Authors: Jeeshma Nambidi Parambath, Gayathri Valsala, Karthik Menon and Shiburaj Sugathan, date: 06/20/2018, view: 357, Q&A: 0

Small heat shock proteins (sHSP) are stress proteins which are ubiquitously found in almost all living organisms. They function as molecular chaperones, which assist in protein folding during translation and in the prevention of irreversible protein ...

Total RNA Extraction from Dinoflagellate Symbiodinium Cells

Author: Tingting Xiang, date: 06/05/2018, view: 332, Q&A: 0

Dinoflagellates are unicellular algae that can have photosynthetic or nonphotosynthetic lifestyles. Dinoflagellates in the genus Symbiodinium can enter endosymbiotic associations with corals, providing the metabolic basis for the highly ...

Root Gall Formation, Resting Spore Isolation and High Molecular Weight DNA Extraction of Plasmodiophora brassicae

Authors: Sara Mehrabi, Suzana Stjelja and Christina Dixelius, date: 06/05/2018, view: 449, Q&A: 0

Isolation of DNA from obligate biotrophic soil-borne plant pathogens is challenging. This is because of their strict requirement of living plant tissue for their growth and propagation. A soil habitat further imposes risk of contamination from other ...

Growing and Pollinating Maize

Authors: Sarah Hake and China Lunde, date: 05/05/2018, view: 363, Q&A: 0

[Abstract] Corn, also known as maize, has been a model organism for genetics since the 1900’s due to the ease of pollinations, large ...

Nuclear Transformation of Chlamydomonas reinhardtii by Electroporation

Author: Tyler M Wittkopp, date: 05/05/2018, view: 690, Q&A: 0

The unicellular green alga Chlamydomonas reinhardtii is an important model organism for studying photosynthesis, acclimation to abiotic stress, cilia biology, and many other biological processes. Many molecular biology tools exist for ...