Is there any mouse Cell line producing CCL20/MIP3alpha?

07 August 2016 8,818 3 View

Can any one suggest protein linkers?

Can anyone suggest a eukaryotic linker for proteins with the property of auto cleavage or protease sensitive and size 3-6 amino acids.

03 April 2016 1,887 3 View

Why recombinant gene is not getting expressed?

I am trying to express fusion peptides of mycobacterium tuberculosis in BL21DE3 and Rosseta DE3 but not getting any expression. Vector I have used is pET23a and pET28b. Gene is fusion of 6...

02 March 2016 7,294 5 View

Is it possible to clone Promoter and RBS in to an expression vector?

I want to know that is there any possibility to clone complete promoter and ribosome binding site in a expression vector to increase the cloning site. I want to clone more than one gene to express...

10 November 2014 8,021 2 View

Will the addition of OADC instead of ADC in 7H9 broth affect the mycobacterial growth?

I would like to know whether the addition of OADC instead of ADC will impact the growth of mycobacterium.

08 September 2014 5,272 4 View

How can I carry out co-expression of mammalian gene into mycobacterium?

I want to express two mammalian gene into single plasmid of mycobacterium. In which way should these gene products should get secreted and separated? So which kind of strategy I should follow and...

07 August 2014 746 3 View

Restriction Ligation

I am trying to ligate my plasmid digested with BamH l and Hind lll and same to my gene of interest but I am unable to ligate them. I have used alkaline phosphatase digestion for plasmid but still...

11 December 2013 9,724 13 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View