Abire Sigbessia

11 Questions 20 Answers 0 Followers

Questions related from Abire Sigbessia

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...

09 September 2019 6,566 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...

09 September 2019 3,756 4 View

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?

09 September 2019 8,314 3 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...

06 June 2019 4,300 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 ?

06 June 2019 9,239 3 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...

04 April 2019 9,418 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?

04 April 2019 2,660 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?

04 April 2019 7,605 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...

03 March 2019 4,153 9 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...

03 March 2019 6,207 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...

03 March 2019 5,708 2 View