Does anyone know a vector which can express protein easily?

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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

Why has the protein loss it's red color?

i'm doing a protein purification that i incorporated with mcherry because i want to visualize that after purification.i've tried low temperature 14 degree as well as 37 degree but the protein and...

08 September 2019 3,682 4 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

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

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

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

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 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 get the vector of a specific vibration mode in phonon dispersion?

For example, I have this phonon dispersion(Fig. 1), and now I would like get vector of atoms which describes TA1 in P. I know v_sim can show vibrational animation of specific mode, but I do not...

20 February 2021 4,332 4 View

Specify the Burgers vectors for dislocations, which cannot be uniquely identified.?

1. A bcc metal single crystal was sliced along (1 -1 0) plane for TEM study. Assuming that the TEM available is equipped with a goniometer stage which tilts up to 45o. The Burgers vector of...

20 February 2021 2,370 1 View

I am working on image transmission, i converted the image into binary vector, i want to extract the MSB of each pixel ?

i have a vector 1*512 size, i want to take the first value every 8 elements and put these elements at the end of the vector,how to do this on matlab ?

19 February 2021 2,352 5 View

Why are Sanger Sequencing reads interrupted in some edited clones?

I have a Crispr/Cas9 edited bulk population and I can briefly check the indel ratios in the bulk population using Sanger sequencing. After cloning single cells, I wanted to check the indel types...

18 February 2021 8,198 6 View

Hi, can anybody help me with the suggestions how can I amplify a stress induced gene of 1107 bp?

I am trying to amplify the TGA1 (Transcription factor 1 ) of Arabidopsis ,which is 1107 bp. I have treated the plants with Salycylic solution over night and extracted the RNA and make cDNA. I...

17 February 2021 7,802 2 View