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Questions related from Ryan Coleman
I am using the clontech adenoX in fusion (system 3) to generate an shRNA adenovirus due to the apparent simplicity of the protocol (no shuttle vector cloning required). For this approach, you...
04 April 2017 5,476 0 View
I have been working on a ChIP for a while, and am not sure what I should be expecting for % Input values of ChIP with specific antibody vs. no antibody or IgG. My protein of interest is not a...
12 December 2015 398 3 View
I have hit a strange roadblock with my experiments. I am doing a ChIP experiment with an HA-tagged protein in H9c2 cells. I used IgG as an isotype control for my antibody, and after isolating DNA...
10 October 2015 7,291 5 View
I am starting a new project looking to see if candidate microRNAs from a bioinformatics scan regulate the gene I study. First I need to see if any of the candidate miRs are differentially...
09 September 2015 977 8 View
I am attempting to determine whether microrna-25 regulates the gene I'm studying. My plan is to transfect a luciferase construct with the 3'UTR of my gene and cotransfect a plasmid with mir-25 to...
09 September 2015 3,429 3 View
I've been working on trying to get a ChIP assay to work on a protein that isn't known to be a transcription factor, but that our lab suspects acts as one in a certain disease state. My project is...
09 September 2015 7,917 14 View
I am starting to work with H9c2 cells. I will be attempting to transfect them with a tagged protein I want to study. I tried a pilot transfection, but when I harvested the cells in about 100 uL of...
08 August 2015 2,509 3 View
I've been trying to develop my technique as a graduate student and am attempting to partition the antifungal drug Nystatin into a liposome preparation consisting of 80% POPC/ 20% cholesterol. I've...
12 December 2014 6,092 4 View
