Martin Chenal

10 Questions 63 Answers 0 Followers

Questions related from Martin Chenal

Can a 5'biotinylated PCR product be used in a PCR cloning kit ?

PCR cloning kits allow to clone a PCR product in a variety of vectors through a simple blunt-ended ligation. However, would this still work with PCR products containing modified ends (obtained...

22 October 2020 4,969 5 View

Pre-running TBE-Urea gels or not, that is the question.?

I am using TBE-Urea gels in PAGE to analyze ssDNA. About half of the protocols (like Qiagen and Biorad) out there insist on pre-running the gel at constant wattage to pre-heat it. However, others...

05 August 2020 4,820 2 View

Sticky pads in orbital shaker : maximum flask volume ?

Title says it all. I need to incubate at 250 rpm big volumes of bacterial cultures in an orbital shaker that only has "sticky pads". I am using 2L erlenmeyers flasks (regular). What is the maximum...

24 July 2020 2,963 0 View

Protein lost during concentration in Amicon device ?

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...

14 February 2020 3,915 4 View

Program to predict restriction sites of novel nuclease ?

I am looking for a program/online tool that can predict the restriction site of novel endonucleases/restriction enzymes based on the pattern of digestion of several DNA substrates like lambda DNA...

06 February 2020 2,222 0 View

Can you use an Amicon-Ultra column to remove HIGH MW proteins ?

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...

02 October 2019 769 3 View

Will protease inhibitor PMSF prevent me from later removing 6xHis-tag ?

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...

20 September 2019 5,455 4 View

Can you ligate 2 PCR products without using a plasmid vector ?

I want to quickly insert a Kanamycine resistance cassette into a gene of Neisseria dentiae (which is naturally competent). To do so, I am planning to amplify by PCR the region containing my gene...

25 June 2019 2,765 3 View

Can you use Tris-glycine PAGE for DNA (gel shift assay/EMSA) ?

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...

22 February 2019 8,826 15 View

How can the complementation of a KO gene enhance the KO phenotype?

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...

11 January 2019 7,748 1 View