Shridhar Kinkar

4 Questions 29 Answers 0 Followers

Questions related from Shridhar Kinkar

What is the minimum time required for a standard ligation reaction using T4 DNA ligase at 1U/ul concentration?

Hello everyone, I have a vector backbone of 5.5Kb in which I need to ligate an insert of ~600bp. The Roche T4 DNA ligase manual I'm using says ligation should be kept for 16h @ 16 deg. Celsius. My...

18 December 2019 4,648 8 View

Is it possible for an E. coli cell to have different morphology during growth?

I have been growing E. coli host in SM6 medium for fermentation. During the run, I did microscopy and found long filamentous rods, which are Gram negative. I have started searching literature for...

14 May 2019 977 8 View

Need to know if higher IPTG concentrations make E. coli expression host undergo a metabolic stress?

Hi, I have been titrating best IPTG dose for highest expression of my GOI. However, I found that after a certain concentration (2.5mM), post-induction OD600 of culture shows lesser growth and as a...

29 January 2018 1,630 3 View

What is the best way to purify tagless and soluble TNFa?

I expressed TNFa in E. coli BL21 (DE3) host and got satisfactory expression. The protein is tagless. My question is how do I purify it? Which chromatography techniques can I use and what should be...

23 September 2014 1,016 17 View