I wonder...
What happen when express rare codon in E. coli?
and Can be express full length protein even contain rare codon? ^-^
31 December 2015 2,907 3 View
My protein is expressed at 20C for 18h with 0.1mM IPTG Anyone can suggest optimal OD for induction?
10 November 2015 3,899 6 View
Which vectors are effective for secreted proteins from E. coli cell into medium?
10 November 2015 2,717 1 View
What is the optimal IPTG concentration and incubation time after induction? Normally, My protein were induced with 0.5mM at 25'C for O/N but now I have limitation about shaker, so i'll to try new...
08 September 2015 1,054 4 View
06 July 2015 9,649 5 View
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
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
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
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
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
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
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
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
I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...
27 February 2021 356 6 View
I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...
27 February 2021 547 3 View