Naghmeh Azadfar

11 Questions 24 Answers 0 Followers

Questions related from Naghmeh Azadfar

How can I stop DNA annealing reaction?

I am annealing 37 DNA oligos together but i would like to stop it in certain time. i tried to add loading dye to stop it but it did not worked. Do you know how can i stop it?

15 December 2015 7,789 5 View

Why dose not a His-tag fused protein eluate during eluation step in FPLC even with > 2 M Imidazole concentration?

I am purifying a His-tag fused protein by Äkta instrument from GE Healthcare. The column in 1 ml HisTrap column. after loading the lysis to the column and washing by washing buffer, i elute the...

03 July 2015 8,678 12 View

What would be the reason for lower extinction coefficient of fluorescent protein than the expected one?

I purified a mRuby2 protein from Bactria. the extinction coefficient of this protein is 113000 M-1cm-1. but my expressed mRuby2 protein has a extinction coefficient of 45000 M-1cm-1. what would be...

09 March 2015 6,597 12 View

How can I express a gene in pBAD vector?

I have a fluorescent protein in a pBAD vector and I want to express and purify it. At first, I cultured a colony in 100 ml LB medium at 37 °C for over night. Then, I inoculated 25 ml from over...

20 February 2015 5,757 9 View

Why is the speed of scaffold lower than assembled DNA origami (6HB) in na agarose gel?

I assembled a DNA origami, a 6HB, and I run a scaffold and assembled 6HB in a agarose gel. In a gel, the band of scaffold is higher than 6HB band. actually, the molecular weight of 6HB is more...

28 August 2014 1,175 2 View

Has anyone used Image J or Fiji to analyze TIRF image stacks of single molecule photobleaching?

I want to look at the step-wise bleaching of membrane proteins tagged with a fluorescent protein. I would need to first identify fluorescent spots on the first frame and then look at the...

01 July 2014 7,607 3 View

When K^2 is not equal to 2/3, how does FRET efficiency change?

I have a labeled protein that the dye does not rotate freely, I measured it by anisotropy method. Now, how can I calculate the correct value for K^2 from anisotropy? This value has an affect on...

14 August 2013 9,563 2 View

How to optimize conditions for labeling of a hydrophobic peptide with Alexa Fluor 647-maleimide dye and Alexa Fluor 546 NHS ester?

Hydrophobic peptide (53%), 15 a.a.-long contains a single Cys-residue and Amine free, can be dissolved in DMF. I need to attach Alexa Fluor647-maleimide to the thiol group of the peptide and Alexa...

25 July 2013 8,893 7 View

Anyone working with CLC Main Workbench program and knows how to add an annotation?

I have a sequence of my construct in CLC Sequence Viewer and I want to change the name of annotation and add another annotation to the construct sequence. Does anyone know how can I do...

12 July 2013 3,755 1 View

Easy and efficient protocol for lysis of spheroplast yeast cell?

I transformed my construct to yeast cells then i labeled my construct inside the cells. Now, I want to prepare lysate from yeast cells to extract the labeled proteins. I prepared spheroplast by...

12 July 2013 7,398 7 View

How can I distinguish between peak-zero FRET and a real low FRET population in an sm-FRET histogram?

I am using an ALEX system to filter direct excitation and leakage of donor dye in red channel and it is possible to select the FRET populations by filtering. But still I have some bursts in the...

10 April 2013 967 7 View