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Riddhi Shah is currently Postdoctoral Research Fellow of the American Heart Association and the research is conducted in Columbia University Medical Center,USA. She has a biological research experience of Columbia University Medical Center, Department of Medicine, New York 09/12/2016 -06/30/2017 Postdoctoral Research: The effects of Sleep Restriction on Endothelial Activation in Pre- and Post-Menopausal Women • Assessing in endothelial cell harvesting biopsy and blood sample collection. • Isolation of circulating endothelial progenitor cells (EPC) and endothelial apoptotic microparticles (EMP). • Assessing EPC and EMP levels by flow cytometry • Isolating platelets and investigating for platelet activation and aggregation by flow cytometry. • White blood cell (WBC) isolation • HUVEC cell culturing and passaging. • Conducting hypoxia experiments on HUVEC cells
• Conducted PCR mutagenesis (Site-specific and random) on iCre recombinase gene cassette followed by cloning into the expression vector. • Tested the expression vector activity with reporter plasmids by Bacterial Solid and Liquid Culture Transformation. • Analyzed variants with routine and gradient PCR reactions (Taq, Q5, KOD, Pfu). • Constructed the TALE – DNA Binding Domain. • Modified the recombinase variants by binding them to TALE to increase their efficiency. • Constructed reporter CHO model cell line to test recombinase expression vectors. • Optimized transfection protocols in CHO and HEK 293 mammalian cells. • Confirmed the activity of the variants in CHO and 293HEK cell lines with transfection reagents. ((Fugene, DNA-In, Lipofectamine, Polyfect, K2-Reagent) • Optimized reportor and expression plasmid concentrations for efficient recombination and integration activities in CHO and 293HEK cells. • Performed Genomic and Total DNA isolation to test the integration and replacement of genes in the genome. Further confirmed using Southern Blotting. • Tested the CRISPR/gRNA setup for Flp variants in 293HEK cells to observe integration activity.
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