How can I express a premiRNA alongside GFP protein?

suppose I have a piggyback vector with this sequence: (5'-pB--promoter--chimeric intron--GFP-?-polyA--pB-3') now I want to express a premiRNA gene beside the gfp protein with same promoter and...

31 December 2017 7,457 7 View

Is there any online tool to compare two amino acid sequences folding?

is there any online tool to compare two amino acid sequences folding with ten amino acids conversion between them?

10 November 2014 5,181 0 View

What's the reason of antibiotic resistance when the plasmid appears to not exist?

I transformed agrobactrium (LBA 4404) with a binary vector with a kanamycin resistance gene. After transformation the colonies grow on the plate with kanamycin (final concentration 50ug/ml), But...

04 May 2013 6,813 15 View

Why the color of precipitated bacteria is black?

When I precipitate the bacteria for making the competent cells, the bacterial pellets are somewhat black. I use LB medium(10+10+5) & DH5-alpha for precipitation. I centrifuge them at 8000 rpm for...

02 March 2013 7,310 5 View

Problems cloning a 300 nucleotide fragment with XhoI and EcoRI restriction sites in pHannibal vector.

I want to clone a 300 nucleotide fragment in pHannibal vector with xhoi and EcoRI restriction sites. I designed two primers with these restriction sites in 5' ends of my...

01 February 2013 7,840 13 View

Does anybody have whole sequence information of binary vector pART27?

Does anybody have or know where to find the whole sequence information of binary vector pART27 [which was constructed by Dr. Andrew Gleave(Plant Molecular Biology 20:1203-1207, 1992)]

01 February 2013 6,273 6 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 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

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 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

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View