Mangesh Kshirsagar

9 Questions 24 Answers 0 Followers

Questions related from Mangesh Kshirsagar

Why did clumps formation observed?

While preparing agarose gel, if we add Agarose powder in already boiled 1X TAE buffer, clumps (precipitation) formation observed. Agarose was not getting dissolved uniformly even after mixing and...

02 February 2017 9,280 2 View

Why plasmid after scale up not same of original?

I have cloned gene in one plasmid and sequence confirmed with Miniprep. I want to scale up same plasmid, for that I transformed my Final Miniprep plasmid into DH5 alpha cells and further used a...

12 December 2016 4,610 5 View

How to religate double digested plasmid?

Hi all, I have XbaI/BamHI digested linearized plasmid. I want to re ligate the same plasmid. Please provide suggestions.

07 July 2016 3,811 5 View

What is the highet transformation efficiency of DH5 alpha (E.coli )get to anyone ever?

If the number of colonies are very high (even hard to count on plate) when transformation done with 1 ng of DNA then how to calculate transformation efficiency? Anyone experienced that even with...

06 June 2016 5,115 4 View

Anybody have done the research related to stability of plasmid DNA at 37°C?

Please share your experience related to the stability of plasmid DNA at 37°C with respect to time (Hrs) in restriction digestion reaction? What is maximum time (Hrs) can we continue restriction...

06 June 2016 8,275 5 View

Why the plasmid not linearizing with NruI restriction enzyme?

I am trying to linearize one circular plasmid with NEB NruI restriction enzyme. I have sequenced my plasmid and confirmed the NruI restriction site. I am using NEB buffer 3.1 in the NruI digestion...

06 June 2016 5,367 5 View

What will be the qualitative and quantitative difference in plasmid scaled up from DH5 alpha (E.coli) and Stbl2 (E.coli)?

If the same plasmid with same insert is used for transformation into DH5 alpha (E.coli) and Stbl2 (E.coli) and if further done scale up. What will be the qualitative and quantitative difference...

06 June 2016 990 5 View

What are the important precautions to get minimum amount of endotoxin during plasmid isolation?

What are the probable reasons by which there are chances of introducing endotoxin during plasmid isolation? How much chances are there to introduce endotoxin through air?

06 June 2016 9,604 4 View

What is the alternative for factor Xa to cleave the MBP tag from proteins expressed in pMAL system?

I am getting nonspecific bands on SDS PAGE after digestion with factor Xa. Is there any alternate cost effective method/component for factor Xa?

08 August 2015 4,884 4 View