Mangal Singh

14 Questions 223 Answers 0 Followers

Questions related from Mangal Singh

How to license a preprint?

I want to submit a research article on bioRxiv, but not sure of the license to use that will be compatible with most of the journals. CC-BY, CC-BY-NC, CC-BY-ND, CC-BY-NC-ND, CC0, or no reuse

11 November 2018 7,153 0 View

Why to use a colorimetric assay for DNA and protein estimation when these molecules give direct absorption at 260 and 280 nm?

All the labs working on DNA and protein have UV/VIS spectrophotometer.

09 September 2017 2,176 3 View

How to go with IMAC fractions eluted in 8M urea?

I am going to perform peptide purification for the first time in denaturing conditions with urea. This will be followed by HPLC for desalting. So my concern is where to store the fractions and for...

08 August 2017 4,715 1 View

What could be the reason for change in RT of the same sample?

I have run 300 uM of tryptophan in isocratic 50% methanol with 40 mM sodium citrate buffer pH 2.25, 400 uM SDS in a 40 min run at 35 degree and absorbance at 280 nm. During two reps the RT changed...

03 March 2017 4,144 8 View

What is the use of 254 nm wavelength in AKTA prime plus purification system?

I was just wondering why besides the 280 nm wavelength there is an option for 254 nm in the Akta Prime plus protein purification system. From a google search I only got to know that the absorbance...

11 November 2016 6,334 2 View

Do we have any agency like OSHA in India?

I was going through online lab safety training provided by NIH and could learn that me and many around me are working in unsafe conditions. Some labs are completely closed without a fume hood or...

06 June 2016 7,814 0 View

What to do with HPLC waste solvents?

We have 60 litres of organic solvents as HPLC waste collected in the solvent bottles, how can these be discarded?

04 April 2016 9,895 5 View

How to store IPTG induced cell pellet for whole cells activity analysis without losing activity?

Currently I am over-expressing the cloned gene in E.coli BL21(DE3) and harvest it and wash it with phosphate buffer. Then making a 20% cell suspension in 50 mM phosphate buffer pH 7.5 containing...

01 January 2016 5,764 4 View

How can I choose HPLC column for lantibiotic purification?

The lantibiotics that are currently purified in the literature, use all different columns like C4, C12 and C18. How to decide which one to use.

10 October 2015 2,367 4 View

What is the amount of DNAse I required to be added in lysozyme lysis of recombinant E. coli cells?

I am breaking open my recombinant e.coli cells (20% solution in tris sucrose buffer) by lysozyme treatment. After the treatment, the solution turns very viscous. Then I centrifuge the solution at...

07 July 2015 6,014 6 View

What components need to be removed from an IMAC purified protein?

My protein was present in 50 mM Sodium Phosphate, 500 mM NaCl and 10 mM Imidazole. It was loaded onto Ni-NTA and washed with same buffer but 20 mM imidazole. Then the protein was eluted using 50...

06 June 2015 3,537 8 View

Do you have knowledge on expressing a protein with high Protein Instability Index?

I have to clone and overexpress a gene, upon analysis of the encoded protein sequence with Expasy ProtParam tool, it is showing a PII of >40, saying that the protein is unstable. How...

07 July 2014 5,125 4 View

How to calculate the flow rate for sephadex G-50 superfine packed gel filtration column using Darcy's law?

I have packed a column with ~50 mL (height=28.5 cm) sephadex G-50 superfine, as required for the high resolution chromatography of a ~9.5 kDa peptide, in a BIORAD econo-column (d=1.5 cm, h=30 cm,...

01 January 1970 2,597 4 View

Can PD-10 column be used for efficient desalting of a 9 kDa peptide?

Its near the exclusion limit of 5 kDa of PD10

01 January 1970 8,085 4 View