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Institute of Life Sciences

3 ARTICLES PUBLISHED IN JoVE

Biochemistry

In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays

Ruchir C. Bobde^*1,2, Ketul Saharan^*1,2, Somanath Baral^1,3, Surajit Gandhi^1,2, Archana Samal^1,2, Rajivgandhi Sundaram¹, Ashish Kumar^1,4, Ajit K. Singh^1,5, Aritreyee Datta¹, Dileep Vasudevan¹

¹Institute of Life Sciences, ²Regional Centre for Biotechnology, ³School of Biotechnology, KIIT University, ⁴Department of Molecular Biophysics and Biochemistry, Yale University, ⁵Department of Pharmacology, University of Vermont College of Medicine

This protocol describes a battery of methods that includes analytical size-exclusion chromatography to study histone chaperone oligomerization and stability, pull-down assay to unravel histone chaperone-histone interactions, AUC to analyze the stoichiometry of the protein complexes, and histone chaperoning assay to functionally characterize a putative histone chaperone in vitro.

Cancer Research

Zebrafish Larvae as a Model to Evaluate Potential Radiosensitizers or Protectors

Amlan P. Mohapatra^*1,3, Deepti Parida^*1,3, Debasish Mohapatra^1,4, Usharani Nayak^2,3, Rajeeb K. Swain², Shantibhusan Senapati¹

¹Tumor microenvironment and animal models lab, Institute of Life Sciences, ²Vascular Biology lab, Institute of Life Sciences, ³Regional Centre for Biotechnology, ⁴School of Biotechnology, KIIT University

The zebrafish has recently been exploited as a model to validate potential radiation modifiers. The present protocol describes the detailed steps to use zebrafish embryos for radiation-based screening experiments and some observational approaches to evaluate the effect of different treatments and radiation.

Biology

Quantification of Circular RNAs Using Digital Droplet PCR

Arundhati Das^*1,2, Debojyoti Das^*1,2, Amaresh C. Panda¹

¹Institute of Life Sciences, ²School of Biotechnology, KIIT University

This protocol describes the detailed method of digital droplet PCR (dd-PCR) for precise quantification of circular RNA (circRNA) levels in cells using divergent primers.