Sanjay Premi

11 Questions 40 Answers 0 Followers

Questions related from Sanjay Premi

How to calculate Synergy for 3 drugs and two timepoints?

Hi all, I have one cell type treated with 3 drugs, 4 sample in total: Untreated, Drug 1, Drug 2, and (Drug 1+2). There are two time points post treatment. The aim of the experiment is to prove...

13 April 2021 837 0 View

How to identify nitrated and nitrosylated proteins in cells/cell lyates?

I aim to detect and identify nitrated and nitrosylated proteins in a particular cell type. These modifications, especially nitrosylation, is highly reversible. I have following 3 questions: 1....

10 March 2018 4,176 4 View

How to prevent DNA loss during purification?

My experiment is to make dsDNA from a pool of variable size fragments (~200 bp to 25 kb). I am using degenerate decamers for second strand synthesis with Phi 29 polymerase. Next step is to purify...

10 January 2016 3,107 4 View

How can I extract biotinylated DNA from Agarose Gel?

Hi, I am trying to remove unligated, biotinylated linkers/adapters from my DNA by running a low melting point agarose gel (0.8%). After cutting the DNA out, and leaving the unligated small sized...

01 July 2015 9,104 4 View

How can I design a random nucleotide sequence so that the random decamers are spaced ~1 to 1.5 kb apart?

I want to make double stranded DNA out of single strands using random decamers or other lengths of random oligos. The conditions is that the random oligos should space apart about 1 kb to 1.5 kb...

18 May 2015 4,953 6 View

Can DMSO in the PCR make amplicon size shift on agarose gel?

I expect a 70 bp amplicon which shows up only with 2% (or higher) DMSO in the PCR. But with DMSO, the size of this amplicon goes from 70 bp to ~110b bp on 3% agarose gel. Same happens when a...

03 October 2014 506 4 View

Is there a good protocol for making blunt ends out of denatured DNA using random hexamers and Klenow (Large Fragment) from NEB?

I am trying to make blunt ends out of nicked plasmid DNA using random hexamers and Klenow polymerase (large fragment) from NEB which has 3'-5' exo but no 5'-3' exonuclease activity. My goal later...

03 September 2014 3,898 4 View

How can I detect carbonyl adducts on proteins?

I have a system that generates carbonyls and dicarbonyls. I am trying to see what effects these carbonyls will have on cellular proteins. I am not looking for common AGEs, HNE or other chemicals...

15 August 2014 1,703 11 View

T4 Endonuclease V is not cutting the UV exposed plasmid - any thoughts?

I am trying to cut ~ 5ug of pUC19 plasmid, exposed to very high UVC dose (~800 J/m2) using T4 endonuclease V from NEB. Everything is being done according to NEB Protocol. I tried increasing the...

11 June 2014 4,027 5 View

What might be causing a very high 260/280 ratio after Phenol:Chloroform extraction of DNA?

I am getting very high 260/280 ratios after a phenol chloroform extraction of small sized DNA fragments. Extra care was taken not to leave any phenol or chloroform in the DNA solution before...

12 May 2014 504 6 View

Any advice on gene expression suppressors in FBS?

I am culturing two different cell types, one with FBS and one without FBS (with horse serum). Is it possible that FBS has suppressors for iNOS or NOX enzymes, or that the FBS acts as an antioxidant?

15 December 2013 1,344 1 View