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Questions related from Martin Chenal
After successful purification of my protein of interest by affinity chromatography (first picture, right side), yielding 50ml of 0.4 mg/ml (estimated by nanodrop), I went on to perform...
02 February 2020 9,627 4 View
Looking at other questions and answers regarding typical cloning procedures (restriction-ligation), I notice almost everyone suggests to GEL purify DNA at several steps. For example ; after a PCR...
12 December 2019 7,770 7 View
I was wondering if one can use an Amicon column to get rid of proteins above the MWCO, by collecting the filtrate instead of the retentate. My protein of interest from affinity chromatography...
10 October 2019 2,993 3 View
I have successfully purified my 13 kDa protein of interest using an Ni-NTA chromatography (see image attached which are my elution fractions). I want to further purify and concentrate it. It...
10 October 2019 5,493 10 View
I am planning on purifying a small 12 kDA protein with a 6x-His tag using Ni-NTA resin. Many protocols recommand the addition of protease inhibitors like PMSF for optimal lysis and purification of...
09 September 2019 536 5 View
I need IPTG to induce big volumes of E.coli cultures to do protein purification. Many suppliers advertise their IPTG as being "dioxane-free". What difference does it make ?
09 September 2019 6,569 4 View
I need to perform an "impossible cloning" as I like to call it, which is inserting a plasmid DNA fragment inside a vector when the restriction sites are not compatible in any way. Basically, I...
06 June 2019 2,199 2 View
Title says it all. I have never published yet, and I was wondering which data is presented in article figures. In my case, I have performed an experiment in biological triplicates at once, and I...
03 March 2019 2,809 3 View
I am planning on doing Electrophoretic Mobility Shift Assay (EMSA), or gel shift assay, to check the binding of a purified protein to different DNA templates. Based on a publication that have...
02 February 2019 9,961 16 View
Hello, I have read on Thermo Fisher's website that EMSA can be done without any kind of labelling, just by post-staining the gel with a fluorescent DNA dye. However, information about this is...
01 January 2019 335 4 View
We think that knocking out a specific gene in Neisseria meningitidis seems to increase recombination events (transformation efficiency, see graph 1). In the overexpressed strain, the...
01 January 2019 4,667 1 View
Hello ! I have linearized a plasmid overnight using a single-cutting restriction enzyme (SalI). I then inactivated the enzyme at 70C for 20 minutes, and stored the linearized plasmid at -20C. I...
01 January 2019 7,822 5 View
