13 Questions 16 Answers 0 Followers
Questions related from Ikram Ullah
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
01 August 2024 9,850 4 View
I am planning to create xenografts of solid tumor cells (sarcoma) in NOD/SCID mice. Our lab has previously established such tumors in these mice but with a certain cell type we have not been able...
18 March 2024 2,926 3 View
Dear All What is the best way to calculate kd (Dissociation constant) for Ligand - Protein binding interactions on Gel using GraphPad Prism. I have an RNA binding protein that I test for binding...
11 April 2018 7,409 4 View
Dear All, I am in some sort of a conundrum because of the results that I have got recently. My RNA binding protein, as the name, binds to ssRNA very well but also binds to dsRNA with equal...
21 November 2017 7,191 5 View
Dear All I want to label in-vitro transcribed RNA with Cy5 dye on one end, what could be the best possible way to do that. For DNA, I am simply labelling one of my Primer with Cy5. If I label Cy5...
15 March 2017 5,228 2 View
Dear All I need to make single stranded DNA fragments. I see that you can biotynilate one of the strand with a biotynilated primer and then separate this specifically with streptavidin-dyna beads....
02 February 2017 9,337 8 View
Dear All, I have been trying to do ChIP qPCRs in Drosophila S2 cells to analyze the recruitment of a protein on certain genes. The problems that I am facing are- the yield is very low...
18 February 2016 5,358 3 View
Dear All, What should be the Molarity of Primers for a qPCR after ChIP, the final concentration in a 25ul reaction. Some protocols suggest using 10uM stocks of each primer (0.5ul each), while...
09 October 2015 8,871 3 View
Dear All, What is the best way of picking the positive clones while establishing a monoclonal cell line expressing your desired transgene. I am currently doing it with a P-20 pipette under a light...
26 May 2015 515 4 View
Dear All How many nucleotides does the 5' exonuclease remove during Gibson Assembly. I mean I was wandering how does the exonuclease recognize the position where it has to stop removing the...
10 May 2015 7,386 3 View
Dear All How do I decide upon the proportions for Gibson Assembly. I have a linearized (digested) vector 2958 bps (27 ng/ul) in which I want to clone two DNA fragments. One fragment is small 410...
12 March 2015 7,404 4 View
Dear All I am looking for any Drosophila melanogaster (Fruitfly) gene that has no intron and produces a transcript of nearly 2.5 Kb. I need this kind of a gene to compare the activity of a...
10 February 2015 2,767 3 View
Dear All I am going to set up Gibson Assembly reactions. The reaction takes place in isothermal buffer which requires 100mM NAD and I have not been able to find this concentration of NAD. I would...
20 January 2015 5,858 8 View
