I wonder...
What happen when express rare codon in E. coli?
and Can be express full length protein even contain rare codon? ^-^
I will propagate pET-22b(+) from Novagen in TOP10(invitrogen) and then transform this plasmid to BL21(DE3) for protein expression
31 December 2015 2,980 3 View
My protein is expressed at 20C for 18h with 0.1mM IPTG Anyone can suggest optimal OD for induction?
10 November 2015 3,970 6 View
Which vectors are effective for secreted proteins from E. coli cell into medium?
10 November 2015 2,786 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 1,141 4 View
My protein show correct size but very light on SDS-PAGE. In addition, Other size band is more bulky than my protein. I'm express in BL21(DE3) at 25'C for 18hr after added 0.5mM IPTG because...
06 July 2015 9,742 5 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
My name is Apurva Saoji. I am a Ph.D scholar in Computer engineering in India. I am looking for international expert in reviewing my PhD thesis, "Competitive Optimization Techniques to Minimize...
07 August 2024 4,600 2 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...
02 August 2024 3,987 1 View
I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...
31 July 2024 225 2 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....
29 July 2024 5,007 0 View
I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...
28 July 2024 366 3 View
I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...
28 July 2024 3,178 3 View
I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...
28 July 2024 7,559 1 View