Hadi H Mohammad

9 Questions 25 Answers 0 Followers

Questions related from Hadi H Mohammad

Repeated inserts ligated to my plasmid??

Hi all I ligated my small insert (37bp) into 3.5kb plasmid, I got colonies but when I check the sequence of my insert, I found more than one insert ligated as a repeats, some clone got three...

02 February 2019 4,095 6 View

How to get the exact number of the successful transformant bacteria???

I transformed 4.5kb plasmid with Ampicillin marker into DH5α by electroporation, I need to get the actual number of transformed bacteria on the plate (i.e. one plasmid per bacteria), after...

08 August 2018 5,920 5 View

Anyone know a suicide plasmid that can be used to delete a gene in E.coli strain?

Anyone know a suicide plasmid that can be used to delete a gene in E.coli strain? I need to use a suicide plasmid to delete a gene in E.coli strain. I used one for gene deletion in Pseudomonas but...

12 December 2017 236 1 View

I found a mysterious thing after protein purification anyone can help?

Dear Dave I am working on a protein (around 125KD) and I have got some mysterious things regarding the purification so I need your help. The protein was expressed using pET28a vector in E.coli...

07 July 2015 6,907 18 View

How can I detect and measure the pyrophosphate PPi releasing from enzymatic reaction?

I have purified a protein which suppose to adenylate amino acid consuming ATP and releasing AMP and PPi, so to test the protein activity I have to measure the released PPi in the solution. Can...

09 September 2014 8,849 18 View

How can I add a DNA linker to a vector?

I am going to design two oligonucleotides (the size about 20 base) which can be annealed together forming two sticky ends then will be ligated to the digested vector with the corresponding enzyme....

04 April 2014 7,996 1 View

How can I chose the best pH for a protein solution?

For the protein to be stable and functional, which is the best pH of the buffer?

02 February 2014 5,448 0 View

Can someone help with flag-tag to the C-terminus of the protein?

Does anyone know how to add Flag-tag to the C-terminus of the protein? What is the principle of the procedure? I am planning to transform bacterial gene to mammalian cells and I have to check the...

01 January 2014 2,487 6 View

How much of EDTA-free protease inhibitor should be used when extracting protein?

I am going to extract protein from BL21 but I do not know how much of EDTA-free protease inhibitor (tablet from Roche diagnostic) should I use.

01 January 2014 10,041 1 View