Shridhar Kinkar

5 Questions 31 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 4,770 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 1,050 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 1,716 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 1,088 17 View

Do you have any experience with ligation independent cloning?

Can anyone suggest me if there's any kit available for LIC? If yes, then which is the most recommended kit?

10 July 2014 524 5 View