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Questions related from Florence Mia
We mainly work with primary cells and the only system that has successfully been use for KO or O/E experiments has been the use of lentiviral systems. However, our lab is abandoning the use of...
06 July 2021 7,332 3 View
I am using a lentiviral TET-ON system for conditional overexpression of my gene of interest in primary kidney cancer cells. I use viral load from transfection of HEK293T cells after 48 and 72...
17 November 2020 2,753 1 View
I am trying to express my gene of interest using a Tetracycline inducible vector. Although I need my gene of interest of be off and only be turned on for a short period of time when needed, I only...
08 July 2017 4,190 19 View
I am having some issues with primer optimization for qPCR. Typically, I first test various concentrations of primers to figure out which concentrations gives the best melting curves and the lowest...
18 January 2017 10,126 2 View
I am performing ChIP on a DNA repair factor in budding yeast. We are interested in looking at the enrichment of this factor in WT and some mutant strains. And I am using a strain that can induce a...
31 August 2016 1,971 2 View
I am trying an experiment in which I measure the NHEJ efficiency of a mutant yeast strain and compare to wild-type to analyze whether the mutation affects the NHEJ process. I take a low copy...
17 December 2015 2,762 3 View
I'm trying to measure the activity of a methyltransferase enzyme for histones and would like to be able to quantify the results. I have tried using 3H labeled SAM and measuring the activity by...
17 July 2015 3,157 5 View
What is the difference between major and minor satellite in human? Do either of them contain alpha-satellite DNA? Do they form distinct domains?
08 December 2014 3,232 1 View
I'm trying to find some target genes for my experiment using results of a ChIP-chip published by another group. They used a non-commercial microarray. In their database, they have ID numbers and...
13 November 2014 6,871 2 View
My substrate is biotinylated and I need to immobilize it on streptavidin slides. However, I'm not sure what conc. or how much to use and how long to incubate. By the way, it needs to be kept cold,...
05 November 2014 5,752 2 View
I have biotin-labeled substrates and I want to visualize them under the microscope. I'm thinking of immobilizing streptavidin on slides or coverslips and then incubate them with my substrates. I'm...
30 October 2014 3,629 6 View
Is it because of heterogeneity in the extent or form of glycosylation or does it have another reason?
14 August 2014 1,074 19 View
Most therapeutic strategies use inhibitors to target a specific function. Are there examples where a drug induces upregulation of a gene? I'm specifically interested in examples in fungi.
22 June 2014 3,355 2 View
There are some examples of actin-independent endocytic pathways in mammalian cells. What about fungi? Are there any studies on fungi?
06 June 2014 384 2 View
When designing inhibitors for an enzyme, is its sub-cellular localization an important factor? Does it matter where the enzyme functions (e.g at cell surface vs. intracellular compartments)?...
03 June 2014 5,353 1 View
Are there known pathways to transport proteins from vacuole to the plasma membrane in budding yeast?
27 May 2014 9,600 1 View
I want to know about methods that are more common for measuring the Kd of two proteins in a cell. I'm thinking of quantitative FRET or similar methods? Please provide references of published papers.
08 May 2014 739 2 View
I have a probe tagged with mVenus and would like to use it as a donor in a FRET experiment to detect interaction between the probe and a histone modification. I'm planning on using an antibody to...
18 April 2014 1,942 1 View
I'm doing some IF experiments with HEK293T cells and I'm trying to fix my cells in a way that they don't detach. I did a quick search and some recommend use of paraformaldehyde fixation for HEK293...
14 April 2014 923 12 View
