Mangesh Kshirsagar

6 Questions 12 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...

13 February 2017 9,571 1 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...

20 December 2016 7,088 4 View

What is reason that after transformation bacterial colonies of unequal sizes?

Sometimes, after plating transformed bacterial cells, colonies grow on plate are of different diameters. What are the factors responsible for uneven sizes of colonies on plate?

13 August 2016 1,023 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...

07 June 2016 2,226 4 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...

07 June 2016 6,527 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?

26 August 2015 2,780 3 View