Jyoti Rustagi

6 Questions 17 Answers 0 Followers

Questions related from Jyoti Rustagi

Why does the marker band wash off as well as the protein from nitrocellulose membrane and PVDF after tbst washing?

I am performing western blot and sometimes the marker band and protein wash away after washing with TBST. I am using the same TBST and all the procedures. But I have tried with different PVDF...

04 March 2023 3,740 3 View

What could be the reason for degration of protein?

My protein has no protease (relevant to the E. coli system)degradation site, as I have checked on expasy. I store my protein at 4 degrees as it aggregated at -20 degrees. It is degrading over time...

04 March 2023 1,077 6 View

Does the concentration of a certain buffer affects the purification?

I am using HEPES 20 mM buffer for the purification of my protein. I have tried different NaCl concentrations like 200 mM to 1M, but I did not see any change in the contamination level. I wanted...

25 December 2022 6,047 4 View

Why there is a difference between the expression method in case of Mut+ vs Muts recombinants of pichia?

In the case of Mut+ recombinants, we first grow cells in a small amount of BMGY till the OD reaches 2-6 and then dilute it in BMMY of OD to 1. But in case of Muts we do the opposite, first we grow...

04 June 2022 9,162 8 View

How to measure the size of plaque using ImageJ?

How to measure the diameter of plaque

22 February 2022 3,415 4 View

What colud be the reason for the slow growth of the bacteria after transformation and what could be the reason that ligation is not successful?

I am working with an engineered (Fusion) gene. It was provided in the cloning vector pUC57 by the company after synthesis i.e., a very high copy number vector. Gene is toxic for E. coli as it...

24 December 2021 6,603 6 View