Is there any particular site to insert the fragment in histag vector for further protein purification?

Do we need to place the insert in a specific site in his tagg vector for further protein purification or can we just use any restriction site present in the vector?

08 September 2019 8,236 3 View

Why do i find another protein size on my gel?

I've cloned a gene which size was around 800bp in pet 32 vector next to n-terminal histagg to further purify the protein. when I expressed the protein there was a big band around 50 kda + my...

08 September 2019 6,492 5 View

Wher can be the issue if the cloning doesn't work?

I"ve been doing a cloning about 4 months already. i've done all the controls and I 've used differents vectors and also I've done vector dephosphorylation though I used double restrictions...

05 June 2019 4,240 12 View

Does anyone know a vector which can express protein easily?

does anyone know an histagg vector that I can use to clone a gene easily and further express my gene protein ?

05 June 2019 9,150 3 View

Is there any diifference between self ligated vector and fragment-vector ligated?

after we have done transformation ,is it possible to compare colonies on plates, i mean is it possible to find that fragment-vector ligated are bigger than self-vector ligated?

03 April 2019 2,593 3 View

What is the importance of iptg for protein expression?

sometimes the protein can be expressed even without adding iptg, so is it necessarily to add iptg for protein expression?

03 April 2019 7,377 2 View

Is it possible to do the insert digestion in two steps?

I'm trying to clone my insert in pet 16b with nde1 and xho1 restrictions enzymes but I'm always getting only self ligated vector ,and the vector control also have many colonies than the...

03 April 2019 9,354 3 View

how can i clone my gene in pet 16b cloning for further protein purification?

i'm trying to clone my gene in pet16b to further make histagg protein purification. Where can I do the cloning for a successful cloning and which restriction enzymes to use? I also want to get a...

02 March 2019 6,140 6 View

Does denaturing conditions alter the protein fluorescence?

I want to express and purify a protein that is incorporated with rfp, red fluorescence protein but the protein is in the pellet. i'm wondering if denaturing conditions like urea use couldn't lead...

02 March 2019 5,649 2 View

Can i use these enzymes to digest my vector and fragments and use the same to test the ligation efficiency?

my primers are F:GCATCATATGatgaatgggaccgcaaaccc and R:GCATCTCGAGttagtctttctggtaaggccg,my fragment that I want to clone start by atgaatggga.......,and my vector is pet16b.i used ndeI and XhoI...

02 March 2019 4,088 9 View

Looking for Materials with high thermal expansion coefficient and high temperature resistance?

03 March 2021 8,272 1 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 View

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 View

What is involved in morphological changes of SEM images of a substance?

In the sem analysis of CaCO3 nanaoparticles, the morphology images showed spherical shaped particles but the proposed morphology for these type of nanoparticles is trigonal. I prepared the...

01 March 2021 1,960 4 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

How to accurately quantify chitin?

Hi all, I have been working on the quantification of chitin in insect samples and feed for a while but I can notice that the quantification methods (e.g. exposure to HCl, NaOH, and weighing sample...

01 March 2021 1,843 2 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

Where can I find atmospheric measurement profiles (temperature, density, etc.) between 300-800 km altitude?

I am investigating thermospheric mass density profiles between 300-800 km altitude. I would like to compare the variability with other parameters, including temperature, electron content,...

01 March 2021 5,902 3 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View