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Phase transition modulation and biophysical characterization of biomolecular condensates using microfluidics

Lab Chip. 2022-07; 
Katherine W Y Chan, Maryam Navi, Jennifer Kieda, Thomas Moran, Daniel Hammers, Shaun Lee, Scott S H Tsai
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Proteins, Expression, Isolation and Analysis … A target DNA sequence for Rv1747-His-TEV is cloned into pET-30a (+) in Escherichia coli DE3 (Genscript) with a 1–310 amino acid sequence of transmembrane protein Rv1747. … Get A Quote

摘要

Membraneless organelles (MLOs) formed through liquid-liquid phase separation (LLPS) are becoming increasingly relevant to understanding viral-host interactions, neurodegenerative disease, and cancer. The modulation of LLPS involves many parameters and components. To describe these modulators, typical studies require laborious, manual sample preparation of different concentrations and costly biological reagents. Here, we introduce a minimal reagent, microfluidic platform to systematically generate samples of different concentrations and trigger phase separation. We demonstrate the platform's utility by constructing phase diagrams describing the modulation of LLPS using an aqueous two-phase system (ATPS) and an ... More

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